Substituted tetrahydro-1H-pyrazolo [3,4-c] pyridines, compositions comprising them, and use

ABSTRACT

Substituted tetrahydro-1H-pyrazolo[3,4-c]pyridines, compositions comprising them and use. The present invention relates in particular to novel substituted tetrahydro-1H-pyrazolo[3,4-c]pyridines having therapeutic activity, which can be used in particular in oncology.

This application claims the benefit of priority of French PatentApplication No. 03/08,441, filed Jul. 10, 2003 and French PatentApplication No. 03/08,442, filed Jul. 10, 2003, both of which areincorporated herein by reference in their entirety.

The present invention relates to novel chemical compounds, particularlynovel tetrahydro-1H-pyrazolo[3,4-c]pyridines, compositions comprisingthem, and to their use as medicinal products.

More particularly, the invention relates to noveltetrahydro-1H-pyrazolo-[3,4-c]pyridines exhibiting anticancer activity,and in particular kinase-inhibiting activity, especially Tie2-inhibitingactivity.

Only a few tetrahydro-1H-pyrazolo[3,4-c]pyridines are known.

Thus, WO 02/012442 discloses tetrahydro-1H-pyrazolo[3,4-c]pyridinessubstituted in the 5-position with an optionally substituted aminogroup. These products are useful in the treatment of cancer and of otherdiseases related to cell proliferation.

P. Krogsgaard-Larsen et al. in Eur. J. Med. Chemical—Chim. Ther. (1979),14(2), p. 157–164, discloses two tetrahydro-1H-pyrazolo[3,4-c]pyridinessubstituted in the 3-position with a hydroxyl group.

WO 96/12720 claims tetrahydro-1H-pyrazolo[3,4-c]pyridines substituted inthe 3-position with substituents chosen from H, alkyl, alkylene,cycloalkyl and methylenecycloalkyl, and in the 1- and 6-positions withvaried substituents. These products are described as inhibitors (i) ofphosphodiesterase type IV (PDE-IV), and (ii) of tumour necrosis factor(TNF), and are, as a result, considered to be useful in the treatment ofinflammatory diseases. No example of a compound according to theinvention is disclosed.

Attempts to obtain effective inhibitors of Tie2 have already beensuccessful in the past (in this respect, see, for example, WO 98/02434;WO 98/41525; WO 99/10325; WO 99/17770; WO 99/54286; WO 99/21859; WO99/55335; WO 00/17202; WO 00/17203; WO 00/27822; WO 00/75139; WO01/37835; WO 01/57008; WO 01/72751; WO 02/060382; WO 02/076396; WO02/076463; WO 02/076954; WO 02/076984; WO 02/076985; WO 02/080926; WO03/004488).

However, none of those documents discloses4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridine derivatives as definedbelow, exhibiting activity against kinases, in particular Tie2.

To this effect, the products in accordance with the invention, accordingto its first aspect, satisfy formula (I) below:

and its tautomers, in which:

L is chosen from a bond, CH₂, CO, SO₂, CONH, COO, NHCO, NH, NHSO₂,SO₂NH, NHCONH, CH₂NH and NHCH₂,

X is chosen from a bond, CH₂, CO, SO₂, CONH and COO;

R1 is chosen from OH, H, alkyl, cycloalkyl, heterocyclyl, aryl,heteroaryl, which is optionally substituted, and, when X is a bond, thenR1 may also be halogen;

R2 is H or is chosen from alkyl, alkylene, cycloalkyl, heterocyclyl,aryl, heteroaryl, which is optionally substituted;

the substituents being chosen independently from R3, O—R3, halogen, NO₂,SO₂—R3, CO—R3, SO₂NH—R3, CONH—R3, N—(R3)₂, NHCO—R3, NHSO₂—R3, NHCONH—R3,NHSO₂NH—R3, OCO—R3, COO—R3, OSO₂—R3, SO₂O—R3, OCONH—R3 and OSO₂NH—R3,where each R3 is chosen independently from H, alkyl, cycloalkyl,alkenyl, aryl, heteroaryl, heterocyclyl, which is optionally substitutedwith halogen, aryl, heteroaryl, R4, OR4 or N(R4)₂, each R4 being chosenindependently from H, C₁–C₄ alkyl and halogenated C₁–C₄ alkyl.

Products in accordance with the invention, according to its firstaspect, are more particularly chosen from products of formula (II)below:

and its tautomers, in which:

X is chosen from a bond, CH₂, CO, SO₂, CONH and COO;

R1 is chosen from alkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl,which is optionally substituted;

R2 is H or is chosen from alkyl, alkylene, cycloalkyl, heterocyclyl,aryl, heteroaryl, which is optionally substituted;

the substituents being chosen independently from R3, O—R3, halogen, NO₂,SO₂—R3, CO—R3, SO₂NH—R3, CONH—R3, N—(R3)₂, NHCO—R3, NHSO₂—R3, NHCONH—R3,NHSO₂NH—R3, OCO—R3, COO—R3, OSO₂—R3, SO₂O—R3, OCONH—R3 and OSO₂NH—R3,where each R3 is chosen independently from H, alkyl, cycloalkyl,alkenyl, aryl, heteroaryl, heterocyclyl, which is optionally substitutedwith halogen, aryl, heteroaryl, OR4 or N(R4)₂, where each R4 is chosenindependently from H and C₁–C₄ alkyl.

Products in accordance with the invention, according to its firstaspect, are more particularly chosen from products of formula (III)below:

and its tautomers, in which:

X is chosen from a bond, CH₂, CO, SO₂, CONH and COO;

R1 is chosen from alkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl,which is optionally substituted;

R2 is H or is chosen from alkyl, alkylene, cycloalkyl, heterocyclyl,aryl, heteroaryl, which is optionally substituted;

in which the substituents are chosen independently from R3, O—R3,halogen, NO₂, SO₂—R3, CO—R3, SO₂NH—R3, CONH—R3, N—(R3)₂, NHCO—R3,NHSO₂—R3, NHCONH—R3, NHSO₂NH—R3, OCO —R3, COO—R3, OSO₂—R3, SO₂O—R3,OCONH—R3 and OSO₂NH—R3, in which each R3 is chosen independently from H,alkyl, cycloalkyl, alkenyl, aryl, heteroaryl, heterocyclyl, which isoptionally substituted with halogen, aryl, heteroaryl, OR4 or N(R4)₂,and in which each R4 is chosen independently from H and C₁–C₄ alkyl.

A product in accordance with the invention is advantageously chosen fromthe products according to its first aspect, in which R1 is heteroaryl,which is optionally substituted, in which a preferred heteroaryl ischosen from benzimidazolyl, indolyl, pyrrolyl, optionally substitutedwith halogen, R4 or O—R4.

More particularly, a preferred heteroaryl is chosen frombenzimidazol-2-yl, indol-2-yl, pyrrol-2-yl, optionally substituted withhalogen, R4 or O—R4.

A product in accordance with the invention according to its first aspectadvantageously has a substituent R2 chosen from phenyl, pyridyl,thienyl, C₁–C₄ alkyl, and C₃–C₇ cycloalkyl, which is optionallysubstituted.

X may advantageously be chosen from CO and SO₂.

A product in accordance with the invention according to its first aspectis advantageously chosen from the products of formula (I) in which R1 isH.

A preferred product is advantageously chosen from the products offormula (I) in which R1 is substituted aryl.

According to a first preferred embodiment, a preferred product isadvantageously chosen from the products of formula (I) in which R1-L isR1-NH—CO, and more preferably when R1 is H.

Very preferably, and according to a second preferred embodiment, apreferred product is advantageously chosen from (i) the products offormula (I), or (ii) preferably the products according to the firstpreferred embodiment, in which X is a bond, and in which R2 is chosenfrom substituted aryl and substituted heteroaryl.

According to a third preferred embodiment, a more preferred product ischosen from the products in accordance with the invention according toits second embodiment, in which R2 is chosen from:

-   -   aryl substituted with NHSO₂—R3 or NHCONH—R3, and    -   heteroaryl substituted with NHSO₂—R3 or NHCONH—R3.

Products according to the third preferred embodiment are advantageouslychosen from:

-   -   aryl substituted with NHSO₂—R3 or NHCONH—R3, and    -   heteroaryl substituted with NHSO₂—R3 or NHCONH—R3, in which aryl        is phenyl and in which heteroaryl is chosen from pyridyl and        pyrimidyl.

According to a fourth embodiment, products according to the thirdpreferred embodiment are advantageously chosen from:

-   -   aryl substituted with NHSO₂—R3 or NHCONH—R3, and    -   heteroaryl substituted with NHSO₂—R3 or NHCONH—R3, in which R3        is chosen from substituted aryl and substituted heteroaryl, in        which R3 is advantageously substituted with a substituent        selected from the group consisting of halogen, R4, OR4 and        N(R4)₂, in which each R4 is chosen independently from H, C₁–C₄        alkyl and halogenated C₁–C₄ alkyl.

According to a fifth embodiment, products according to the fourthpreferred embodiment are advantageously chosen from:

A product in accordance with the invention according to its first aspectmay be in:

-   -   1) racemic form, or    -   2) a form enriched in a stereoisomer, or    -   3) a form enriched in an enantiomer;        and may optionally be salified.

According to a second aspect, the invention relates to pharmaceuticalcompositions comprising a product as defined above, in combination witha pharmaceutically acceptable excipient.

According to a third aspect, the invention relates to the use of aproduct as defined above, as an agent for modulating the activity of akinase. A preferred kinase will advantageously be chosen from Tie2 andKDR. Tie2 is more preferred.

According to its third aspect, the invention relates to the use of aproduct as defined above, for producing a medicinal product that isuseful for treating a pathological condition, in particular cancer.

Products in accordance with the invention can be obtained by methodswell known to those skilled in the art, in particular as regards thetechniques of coupling between an acid and an amine; see, for example,J. March, Advanced organic chemistry, (J. Wiley & Sons, ed.), fourthedition, 1992.

The products of the invention are useful as agents which inhibit areaction catalysed by a kinase. Tie2 is a kinase for which the productsof the invention will be particularly useful as inhibitors. Theseproducts can also be used as inhibitors of other kinases, such as KDR.

Reasons for which the kinases are chosen are given below:

Tie2

Tie-2 (TEK) is a member of a family of tyrosine kinase receptorsspecific for endothelial cells. Tie2 is the first receptor with tyrosinekinase activity for which both the agonist (angiopoietin 1 or Ang1),which stimulates autophosphorylation of the receptor and cell signalling[S. Davis et al (1996) Cell 87, 1161–1169] and the antagonist(angiopoietin 2 or Ang2) [P.C. Maisonpierre et al. (1997) Science 277,55–60] are known. Angiopoietin 1 can synergize with VEGF in the finalstages of neoangiogenesis [AsaharaT. Circ. Res.(1998) 233–240]. Knockoutexperiments and transgenic manipulations of the expression of Tie2 or ofAng1 result in animals which exhibit vascularization defects [D. J.Dumont et al (1994) Genes Dev. 8, 1897–1909 and C. Suri (1996) Cell 87,1171–1180]. The binding of Ang1 to its receptor results inautophosphorylation of the kinase domain of Tie2, which is essential forneovascularization and for the recruitment and the interaction of thevessels with pericytes and smooth muscle cells; these phenomenacontribute to the maturing and the stability of the newly formed vessels[P. C. Maisonpierre et al (1997) Science 277, 55–60]. Lin et al (1997),J. Clin. Invest. 100, 8: 2072–2078 and Lin P. (1998) PNAS 95, 8829–8834,have shown inhibition of tumour growth and vascularization and adecrease in lung metastases during adenoviral infections or duringinjections of the Tie-2 (Tek) extracellular domain in breast tumour andmelanoma xenograph models.

Tie2 inhibitors can be used in situations where neovascularization takesplace inappropriately (i.e. in diabetic retinopathy, chronicinflammation, psoriasis, Kaposi's sarcoma, chronic neovascularizationdue to macular degeneration, rheumatoid arthritis, infantilehaemoangioma and cancers).

KDR

KDR (Kinase insert Domain Receptor), also known as VEGF-R2 (VascularEndothelial Growth Factor Receptor 2), is expressed only in endothelialcells. This receptor binds to the angiogenic growth factor VEGF and thusacts as a mediator to a transduction signal via the activation of itsintracellular kinase domain. Direct inhibition of kinase activity ofVEGF-R2 makes it possible to reduce the phenomenon of angiogenesis inthe presence of exogenous VEGF (Vascular Endothelial Growth Factor)(Strawn et al., Cancer Research, 1996, vol. 56, p. 3540–3545). Thisprocess has been demonstrated in particular using VEGF-R2 mutants(Millauer et al., Cancer Research, 1996, vol. 56, p. 1615–1620). TheVEGF-R2 receptor does not appear to have any function in adults otherthan that related to the angiogenic activity of VEGF. Consequently, aselective inhibitor of the kinase activity of VEGF-R2 should only showslight toxicity.

In addition to this central role in the dynamic angiogenic process,recent results suggest that VEGF expression contributes to the survivalof tumour cells after chemotherapy and radiotherapy, emphasizing thepotential synergy of KDR inhibitors with other agents (Lee et al. CancerResearch, 2000, vol. 60, p. 5565–5570).

Experimental Section

Method A: Analysis by LC/MS

The LC/MS analyses were carried out on a Micromass model LCT deviceconnected to an HP 1100 device. The abundance of the products wasmeasured using an HP G1315A diode array detector over a wavelength of200–600 nm and a Sedex 65 light scattering detector. The mass spectrawere acquired over a range of from 180 to 800. The data were analysedusing the Micromass MassLynx software. Separation was carried out on aHypersil BDS C18, 3 μm (50×4.6 mm) column, eluting with a lineargradient of from 5 to 90% of acetonitrile comprising 0.05% (v/v) oftrifluoroacetic acid (TFA) in water comprising 0.05% (v/v) TFA, over 3.5min at a flow rate of 1 ml/min. The total analysis time, including theperiod for re-equilibrating the column, is 7 min.

Method B: Purification by LC/MS:

The products were purified by LC/MS using a Waters FractionsLynx systemcomposed of a Waters model 600 gradient pump, a Waters model 515regeneration pump, a Waters Reagent Manager dilution pump, a Watersmodel 2700 auto-injector, two Rheodyne model LabPro valves, a Watersmodel 996 diode array detector, a Waters model ZMD mass spectrometer anda Gilson model 204 fraction collector. The system was controlled by theWaters FractionLynx software. Separation was carried out alternately ontwo Waters Symmetry columns (C₁₈, 5 μM, 19×50 mm, catalogue reference186000210), one column undergoing regeneration with a 95/5 (v/v)water/acetonitrile mixture comprising 0.07% (v/v) of trifluoroaceticacid, while the other column was being used for separation. The columnswere eluted using a linear gradient of from 5 to 95% of acetonitrilecomprising 0.07% (v/v) of trifluoroacetic acid in water comprising 0.07%(v/v) of trifluoroacetic acid, at a flow rate of 10 ml/min. At theoutlet of the separation column, one-thousandth of the effluent isseparated by means of an LC Packing Accurate, diluted with methylalcohol at a flow rate of 0.5 ml/min and sent to the detectors, in aproportion of 75% to the diode array detector and the remaining 25% tothe mass spectrometer. The rest of the effluent (999/1000) is sent tothe fraction collector, where the flow is discarded for as long as themass of the expected product is not detected by the FractionLynxsoftware. The molecular formulae of the expected products are suppliedto the FractionLynx software, which actuates the collection of theproduct when the mass signal detected corresponds to the ion [M+H]⁺and/or to [M+Na]⁺. In certain cases, depending on the results of theanalytical LC/MS, when an intense ion corresponding to [M+2H]⁺⁺ wasdetected, the value corresponding to half the calculated molecular mass(MW/2) is also supplied to the FractionLynx software. Under theseconditions, the collection is also actuated when the mass signal of theion [M+2H]⁺⁺ and/or [M+Na+H]⁺⁺ are detected.

Method C: EI Analysis

The mass spectra were produced by electron impact (70 eV) on a FinniganSSQ 7000 spectrometer.

Method D: NMR Analysis

The NMR spectra were produced on a Bruker Avance 300 spectrometer and aBruker Avance DRX 400 spectrometer.

tert-Butyl4-(diazoethoxycarbonylmethyl)-4-hydroxypiperidine-1-carboxylate

CAS Name d MM eq mmol g ml 1 79099- N-Boc- 199.25 1.00 50.19 10.00 07-3piper- idinone 2 623-73- Ethyl 1.085 114.1 1.05 52.70 6.01 5.54 4 diazo-acetate 3 109-72- 1.6 M 1.60 80.30 50.19 8 BuLi hexane 4 108-18- Diiso-0.720 101.19 1.60 80.30 8.13 11.29 9 propyl- amine 5 109-99- THf on 10500 9 4′ mole- vol. cular sieve 6 64-19-7 100% 1.050 60.05 5.00 250.9415.07 14.35 AcOH

A freshly prepared solution of LDA (prepared by the dropwise addition,under an inert atmosphere at −78° C., of 50.19 ml of 1.6 M of BuLi inhexane to a solution of 11.29 ml of diisopropylamine in 200 ml of dryTHF) is added dropwise, under an inert atmosphere at −78° C., onto 10.0g of N-Boc-piperidinone in suspension and 5.54 ml of ethyl diazoacetatein 300 ml of dry THF. The mixture is stirred at −78° C. for 4 hours andis then decomposed at −78° C. with 14.35 ml of concentrated AcOH. Themixture obtained is left at ambient temperature overnight, and thesolvent is then evaporated off under reduced pressure, to 1/10 of itsinitial volume, diluted in diisopropyl ether, and washed 4 times with asaturated NaHCO₃ solution. The organic phase is dried over MgSO₄. Thehydrated salt is removed by filtration and the dry filtrate isconcentrated under reduced pressure so as to give 15.12 g of a viscousyellow oil. LC/MS: RT=2.84; [M+1]+=304.33. The product is used as it isfor the subsequent step.

tert-Butyl4-(diazoethoxycarbonylmethyl)-3,6-dihydro-2H-pyridine-1-carboxylate

GAS Name d MM eq mmol g ml 1 P-31391- 313.35  1 48.25 15.12 106-4 210025- POCl₃ 1.67 153.33  2 96.51 14.80 8.86 87-3 3 110-86- Pyridine0.983 79.1 20 965.06 76.34 77.66 1 (4Å m.s.) 4 108-20- iPr₂O  5 250 3vol 5 1310- 0.1 M 40  1 48.25 483 73-2 NaOH

78.0 ml of dry pyridine are added to a solution of 15.12 g of tert-butyl4-(diazoethoxycarbonylmethyl)-4-hydroxypiperidine-1-carboxylate into 250ml of iPr₂O. The mixture is cooled to −10° C. and 8.86 ml of POCl₃ areadded slowly with vigorous stirring. The mixture is then left to returnto ambient temperature for 12 hours with stirring. The reaction mixtureis decomposed with 500 ml of a 0.1M NaOH solution, and is then extracted3 times with EtOAc. The organic phase is washed with a saturated NaClsolution and dried over MgSO₄. The hydrated salt is removed byfiltration and the dry filtrate is concentrated under reduced pressureto 1/10 of its initial volume. LC/MS:RT=4.57; [M+1]+=296.31. The productis used as it is for the subsequent step.

6-tert-Butyl 3-ethyl2,4,5,7-tetrahydropyrazolo[3,4-c]pyridyl-3,6-dicarboxylate

CAS Name d MM eq mmol mg ml 1 P-31391-120-4 285.35 1.0 48.25 2 108-83-3PhMe 150

The solution of tert-butyl4-(diazoethoxycarbonylmethyl)-3,6-dihydro-2H-pyridyl-1-carboxylate inPy/EtOAc obtained in the preceding step is added dropwise to 150 ml oftoluene reflux. The azeotrope Py/PhMe is distilled at a rate equivalentto the rate of addition. One hour after the end of addition, thesolution is left to cool to ambient temperature, the solvent isevaporated off under reduced pressure and the crude product obtained(15.05 g) is purified by flash chromatography (SiO₂, CH₂Cl₂/MeOH 1%NH₃7M_((MeOH)) 40:1 then 30:1 then 20:1). The solvent is evaporated offand 10.05 g (71% over 3 steps) of a black solid are obtained: LC/MS:RT=3.88; [M+1]+=296.27.

(tert-Butyl2,4,5,7-tetrahydropyrazolo[3,4-c]pyridyl-6-carboxylate)-3-carboxylicacid

Cas Name d MM eq mmol g ml 1 P-31391- 295.34 1.0 34.03 10.05 123-1 21310-66- LiOH.H₂O 41.96 1.1 37.43 1.57 3 3 67-56-1 MeOH 10 375 vol. 47732-18- H₂O 1 38 5 vol

1.57 g of LiOH and 38 ml of water are added to a solution of 10.05 g of6-tert-butyl butyl 3-ethyl2,4,5,7-tetrahydropyrazolo[3,4-c]pyridyl-3,6-dicarboxylate in 375 ml ofMeOH. The mixture obtained is refluxed overnight. The solution is cooledto ambient temperature and is then acidified to pH=2 with 50 ml of a 1MHCl solution. The solution is then extracted 4 times with EtOAc. Theorganic phase is washed with a saturated NaCl solution and then driedover Na₂SO₄. The salt obtained is removed by filtration and the solventis evaporated off under reduced pressure so as to produce 8.90 g (98%)of a white solid. LC/MS: RT=3.21; [M+1]+=268.23.

Preparation of a Library of Products:

tert-Butyl 3-(alkylcarbamoyl, arylcarbamoyl, heteroarylcarbamoyl,etc.)-2,4,5,7-tetrahydropyrazolo[3,4-c]pyridine-6-carboxylate

CAS Name d MM mmol g ml 1 P-31391-031-5 267.28 1.0 3.741 1.00 amine 2.06.744 2 538-75-0 DCC 206.33 1.0 3.741 0.772 3 2592-95-2 HOBt.H₂O 153.131.5 5.612 0.859 4 68-12-2 DMF 5   19 vol.General Method:

DCC and HOBT.H₂O in solution in DMF with 2 eq of amine (R, Ar, orHet)-NH₂ are added to a solution of 1 eq of tert-butyl(2,4,5,7-tetrahydro-pyrazolo[3,4-c]pyridyl-6-carboxylate)-3-carboxylicacid in DMF and the mixture is stirred at ambient temperature for 3 h.The solvent is evaporated off under reduced pressure at 35° C.overnight. The crude product obtained is purified by flashchromatography (SiO₂, CH₂Cl₂/MeOH 1% NH₃7M_((MeOH)) 20:1 then 10:1 then5:1, according to the products).

List of amines R1-NH₂ used (Table 1) [Comment: R1-NH₂═(R, Ar, orHet)—NH₂]:

TABLE 1 Reference number of the amine Structure 1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

3-(Alkylcarbamoyl, arylcarbamoyl, heteroarylcarbamoyl,etc.)-4,5,6,7-tetrahydro -2H-pyrazolo[3,4-c]pyridin-6-iumtrifluoroacetate

General Method:

16 eq of tert-butyl 3-(alkylcarbamoyl, arylcarbamoyl,heteroarylcarbamoyl,etc.)-2,4,5,7-tetrahydropyrazolo[3,4-c]pyridine-6-carboxylate in a 1:1THF/water solution, and the solution is refluxed for 2 h. The solvent isevaporated off under reduced pressure and the viscous oil collected isdried under vacuum overnight. The product thus obtained is used withoutpurification, in the subsequent step.

6-(Alkyl, aryl, heteroaryl,etc.)carbonyl-4,5,6,7-tetrahydro-2H-pyrazolo[3,4c]pyridiene-3-(alkyl,aryl, hetyl, etc.)amide

General Method:

A solution of 2.5 M HOBt.H₂O (2 eq) in DMF, a solution of 0.833M HBTU inDMF (2 eq), a solution of 2.5M DIPEA (4 eq) in DMF and a suspension or asolution at an appropriate concentration of an R₂COOH (2 eq) in DMF areadded, in order, to a solution of 1 eq of 3-(alkylcarbamoyl,arylcarbamoyl, heteroarylcarbamoyl,etc.)-4,5,6,7-tetrahydro-2H-pyrazolo[3,4-c]pyridin-6-iumtrifluoroacetate in DMF. The solutions are stirred overnight at ambienttemperature and are then acidified with 100 μl of 100% AcOH, filteredand purified by preparative LC/MS.

List of the acids R2COOH used (Table 2):

TABLE 2 Reference number of the acid Nomenclature 11-Phenyl-1-cyclopropylcarboxylic acid 2 Acetic acid 3 Propiolic acid 4Crotonic acid 5 Vinylacetic acid 6 Pyruvic acid 7 Sarcosine 8Methoxyacetic acid 9 Lactic acid 10 3,3-Dimethylacrylic acid 11Cyclopropylacetic acid 12 Valeric acid 13 N,N-dimethylglycine 143-Mercaptopropionic acid 15 (Methylthio)acetic acid 16Pyrrole-2-carboxylic acid 17 1-Cyanocyclopropanecarboxylic acid 182-Furoic acid 19 4-Pyrazolecarboxylic acid 20 Imidazole-4-carboxylicacid 21 1-Cyclopentenecarboxylic acid 22 Acid 23 Acetoxyacetic acid 24Hydantoic acid 25 Benzoic acid 26 Nicotinic acid 27 2-Pyrazinecarboxylicacid 28 o-Toluic acid 29 Phenylacetic acid 30 Salicylic acid 312-Fluorobenzoic acid 32 3-Cyanobenzoic acid 33 4-Vinylbenzoic acid 342-Phenylpropionic acid 35 N-Methylanthranilic acid 36 2-Methoxybenzoicacid 37 2-Hydroxyphenylacetic acid 38 4-Hydroxymethylbenzoic acid 392-Fluorophenylbenzoic acid 40 2,6-Difluorobenzoic acid 41Indole-3-carboxylic acid 42 3,5-Dimethylphenylacetic acid 433-(Dimethylamino)benzoic acid 44 Piperonylic acid 45 DL-tropic acid 463-Methoxyphenylacetic acid 47 3-Methoxysalicylic acid 484-(Methylthio)benzoic acid 49 2-Chlorophenylacetic acid 50 2-Naphthoicacid 51 2-Chloro-6-fluorobenzoic acid 52 1-Methylindole-3-carboxylicacid 53 3-Acetamidobenzoic acid 54 4-(Dimethylamino)salicylic acid 552,3-Dimethoxybenzoic acid 56 4-Chlorophenylpropionic acid 572-Chloromandelic acid 58 2-Chloro-6-fluorophenylacetic acid 591-Phenyl-1-cyclopentanecarboxylic acid 60 2,6-Dichlorobenzoic acid 613-Methyl-2-phenylvaleric acid 62 4-Phenylbenzoic acid 632-Chloro-4-nitrobenzoic acid 64 2-Benzylbenzoic acid 65 2-Phenoxybenzoicacid 66 2-Ethoxy-1-naphthoic acid 67 4-(4-N-Propylphenyl)benzoic acid 683,5-Dibromosalicylic acid 69 2,6-Dichlorophenylacetic acid 70Cyanoacetic acidResults

The following products are prepared according to the process describedabove.

In order to simplify the representation of the products in Table 3 whichfollows, the pyrazolopiperidine ring presented in Scheme A is symbolizedby the letter H, the amines R1-NH₂ which are linked to H are symbolizedby the letter B followed by a number ranging from 1 to 15, correspondingto the products listed in Table 1, and the acids R2-COOH which arelinked to H are symbolized by the letter A followed by a number from 1to 70, corresponding to the products listed in Table 2.

Thus, a product referred to as A1-H-B1 corresponds to the followingstructure:

TABLE 3

A1-H-B1 A2-H-B1 A3-H-B1 A4-H-B1 A5-H-B1 A6-H-B1 A7-H-B1 A1-H-B2 A2-H-B2A3-H-B2 A4-H-B2 A5-H-B2 A6-H-B2 A7-H-B2 A1-H-B3 A2-H-B3 A3-H-B3 A4-H-B3A5-H-B3 A6-H-B3 A7-H-B3 A1-H-B4 A2-H-B4 A3-H-B4 A4-H-B4 A5-H-B4 A6-H-B4A7-H-B4 A1-H-B5 A2-H-B5 A3-H-B5 A4-H-B5 A5-H-B5 A6-H-B5 A7-H-B5 A1-H-B6A2-H-B6 A3-H-B6 A4-H-B6 A5-H-B6 A6-H-B6 A7-H-B6 A1-H-B7 A2-H-B7 A3-H-B7A4-H-B7 A5-H-B7 A6-H-B7 A7-H-B7 A1-H-B8 A2-H-B8 A3-H-B8 A4-H-B8 A5-H-B8A6-H-B8 A7-H-B8 A1-H-B9 A2-H-B9 A3-H-B9 A4-H-B9 A5-H-B9 A6-H-B9 A7-H-B9A1-H-B10 A2-H-B10 A3-H-B10 A4-H-B10 A5-H-B10 A6-H-B10 A7-H-B10 A1-H-B11A2-H-B11 A3-H-B11 A4-H-B11 A5-H-B11 A6-H-B11 A7-H-B11 A1-H-B12 A2-H-B12A3-H-B12 A4-H-B12 A5-H-B12 A6-H-B12 A7-H-B12 A1-H-B13 A2-H-B13 A3-H-B13A4-H-B13 A5-H-B13 A6-H-B13 A7-H-B13 A1-H-B14 A2-H-B14 A3-H-B14 A4-H-B14A5-H-B14 A6-H-B14 A7-H-B14 A1-H-B15 A2-H-B15 A3-H-B15 A4-H-B15 A5-H-B15A6-H-B15 A7-H-B15 A8-H-B1 A9-H-B1 A10-H-B1 A11-H-B1 A12-H-B1 A13-H-B1A14-H-B1 A8-H-B2 A9-H-B2 A10-H-B2 A11-H-B2 A12-H-B2 A13-H-B2 A14-H-B2A8-H-B3 A9-H-B3 A10-H-B3 A11-H-B3 A12-H-B3 A13-H-B3 A14-H-B3 A8-H-B4A9-H-B4 A10-H-B4 A11-H-B4 A12-H-B4 A13-H-B4 A14-H-B4 A8-H-B5 A9-H-B5A10-H-B5 A11-H-B5 A12-H-B5 A13-H-B5 A14-H-B5 A8-H-B6 A9-H-B6 A10-H-B6A11-H-B6 A12-H-B6 A13-H-B6 A14-H-B6 A8-H-B7 A9-H-B7 A10-H-B7 A11-H-B7A12-H-B7 A13-H-B7 A14-H-B7 A8-H-B8 A9-H-B8 A10-H-B8 A11-H-B8 A12-H-B8A13-H-B8 A14-H-B8 A8-H-B9 A9-H-B9 A10-H-B9 A11-H-B9 A12-H-B9 A13-H-B9A14-H-B9 A8-H-B10 A9-H-B10 A10-H-B10 A11-H-B10 A12-H-B10 A13-H-B10A14-H-B10 A8-H-B11 A9-H-B11 A10-H-B11 A11-H-B11 A12-H-B11 A13-H-B11A14-H-B11 A8-H-B12 A9-H-B12 A10-H-B12 A11-H-B12 A12-H-B12 A13-H-B12A14-H-B12 A8-H-B13 A9-H-B13 A10-H-B13 A11-H-B13 A12-H-B13 A13-H-B13A14-H-B13 A8-H-B14 A9-H-B14 A10-H-B14 A11-H-B14 A12-H-B14 A13-H-B14A14-H-B14 A8-H-B15 A9-H-B15 A10-H-B15 A11-H-B15 A12-H-B15 A13-H-B15A14-H-B15 A15-H-B1 A16-H-B1 A17-H-B1 A18-H-B1 A19-H-B1 A20-H-B1 A21-H-B1A15-H-B2 A16-H-B2 A17-H-B2 A18-H-B2 A19-H-B2 A20-H-B2 A21-H-B2 A15-H-B3A16-H-B3 A17-H-B3 A18-H-B3 A19-H-B3 A20-H-B3 A21-H-B3 A15-H-B4 A16-H-B4A17-H-B4 A18-H-B4 A19-H-B4 A20-H-B4 A21-H-B4 A15-H-B5 A16-H-B5 A17-H-B5A18-H-B5 A19-H-B5 A20-H-B5 A21-H-B5 A15-H-B6 A16-H-B6 A17-H-B6 A18-H-B6A19-H-B6 A20-H-B6 A21-H-B6 A15-H-B7 A16-H-B7 A17-H-B7 A18-H-B7 A19-H-B7A20-H-B7 A21-H-B7 A15-H-B8 A16-H-B8 A17-H-B8 A18-H-B8 A19-H-B8 A20-H-B8A21-H-B8 A15-H-B9 A16-H-B9 A17-H-B9 A18-H-B9 A19-H-B9 A20-H-B9 A21-H-B9A15-H-B10 A16-H-B10 A17-H-B10 A18-H-B10 A19-H-B10 A20-H-B10 A21-H-B10A15-H-B11 A16-H-B11 A17-H-B11 A18-H-B11 A19-H-B11 A20-H-B11 A21-H-B11A15-H-B12 A16-H-B12 A17-H-B12 A18-H-B12 A19-H-B12 A20-H-B12 A21-H-B12A15-H-B13 A16-H-B13 A17-H-B13 A18-H-B13 A19-H-B13 A20-H-B13 A21-H-B13A15-H-B14 A16-H-B14 A17-H-B14 A18-H-B14 A19-H-B14 A20-H-B14 A21-H-B14A15-H-B15 A16-H-B15 A17-H-B15 A18-H-B15 A19-H-B15 A20-H-B15 A21-H-B15A22-H-B1 A23-H-B1 A24-H-B1 A25-H-B1 A26-H-B1 A27-H-B1 A28-H-B1 A22-H-B2A23-H-B2 A24-H-B2 A25-H-B2 A26-H-B2 A27-H-B2 A28-H-B2 A22-H-B3 A23-H-B3A24-H-B3 A25-H-B3 A26-H-B3 A27-H-B3 A28-H-B3 A22-H-B4 A23-H-B4 A24-H-B4A25-H-B4 A26-H-B4 A27-H-B4 A28-H-B4 A22-H-B5 A23-H-B5 A24-H-B5 A25-H-B5A26-H-B5 A27-H-B5 A28-H-B5 A22-H-B6 A23-H-B6 A24-H-B6 A25-H-B6 A26-H-B6A27-H-B6 A28-H-B6 A22-H-B7 A23-H-B7 A24-H-B7 A25-H-B7 A26-H-B7 A27-H-B7A28-H-B7 A22-H-B8 A23-H-B8 A24-H-B8 A25-H-B8 A26-H-B8 A27-H-B8 A28-H-B8A22-H-B9 A23-H-B9 A24-H-B9 A25-H-B9 A26-H-B9 A27-H-B9 A28-H-B9 A22-H-B10A23-H-B10 A24-H-B10 A25-H-B10 A26-H-B10 A27-H-B10 A28-H-B10 A22-H-B11A23-H-B11 A24-H-B11 A25-H-B11 A26-H-B11 A27-H-B11 A28-H-B11 A22-H-B12A23-H-B12 A24-H-B12 A25-H-B12 A26-H-B12 A27-H-B12 A28-H-B12 A22-H-B13A23-H-B13 A24-H-B13 A25-H-B13 A26-H-B13 A27-H-B13 A28-H-B13 A22-H-B14A23-H-B14 A24-H-B14 A25-H-B14 A26-H-B14 A27-H-B14 A28-H-B14 A22-H-B15A23-H-B15 A24-H-B15 A25-H-B15 A26-H-B15 A27-H-B15 A28-H-B15 A29-H-B1A30-H-B1 A31-H-B1 A32-H-B1 A33-H-B1 A34-H-B1 A35-H-B1 A29-H-B2 A30-H-B2A31-H-B2 A32-H-B2 A33-H-B2 A34-H-B2 A35-H-B2 A29-H-B3 A30-H-B3 A31-H-B3A32-H-B3 A33-H-B3 A34-H-B3 A35-H-B3 A29-H-B4 A30-H-B4 A31-H-B4 A32-H-B4A33-H-B4 A34-H-B4 A35-H-B4 A29-H-B5 A30-H-B5 A31-H-B5 A32-H-B5 A33-H-B5A34-H-B5 A35-H-B5 A29-H-B6 A30-H-B6 A31-H-B6 A32-H-B6 A33-H-B6 A34-H-B6A35-H-B6 A29-H-B7 A30-H-B7 A31-H-B7 A32-H-B7 A33-H-B7 A34-H-B7 A35-H-B7A29-H-B8 A30-H-B8 A31-H-B8 A32-H-B8 A33-H-B8 A34-H-B8 A35-H-B8 A29-H-B9A30-H-B9 A31-H-B9 A32-H-B9 A33-H-B9 A34-H-B9 A35-H-B9 A29-H-B10A30-H-B10 A31-H-B10 A32-H-B10 A33-H-B10 A34-H-B10 A35-H-B10 A29-H-B11A30-H-B11 A31-H-B11 A32-H-B11 A33-H-B11 A34-H-B11 A35-H-B11 A29-H-B12A30-H-B12 A31-H-B12 A32-H-B12 A33-H-B12 A34-H-B12 A35-H-B12 A29-H-B13A30-H-B13 A31-H-B13 A32-H-B13 A33-H-B13 A34-H-B13 A35-H-B13 A29-H-B14A30-H-B14 A31-H-B14 A32-H-B14 A33-H-B14 A34-H-B14 A35-H-B14 A29-H-B15A30-H-B15 A31-H-B15 A32-H-B15 A33-H-B15 A34-H-B15 A35-H-B15 A36-H-B1A37-H-B1 A38-H-B1 A39-H-B1 A40-H-B1 A41-H-B1 A42-H-B1 A36-H-B2 A37-H-B2A38-H-B2 A39-H-B2 A40-H-B2 A41-H-B2 A42-H-B2 A36-H-B3 A37-H-B3 A38-H-B3A39-H-B3 A40-H-B3 A41-H-B3 A42-H-B3 A36-H-B4 A37-H-B4 A38-H-B4 A39-H-B4A40-H-B4 A41-H-B4 A42-H-B4 A36-H-B5 A37-H-B5 A38-H-B5 A39-H-B5 A40-H-B5A41-H-B5 A42-H-B5 A36-H-B6 A37-H-B6 A38-H-B6 A39-H-B6 A40-H-B6 A41-H-B6A42-H-B6 A36-H-B7 A37-H-B7 A38-H-B7 A39-H-B7 A40-H-B7 A41-H-B7 A42-H-B7A36-H-B8 A37-H-B8 A38-H-B8 A39-H-B8 A40-H-B8 A41-H-B8 A42-H-B8 A36-H-B9A37-H-B9 A38-H-B9 A39-H-B9 A40-H-B9 A41-H-B9 A42-H-B9 A36-H-B10A37-H-B10 A38-H-B10 A39-H-B10 A40-H-B10 A41-H-B10 A42-H-B10 A36-H-B11A37-H-B11 A38-H-B11 A39-H-B11 A40-H-B11 A41-H-B11 A42-H-B11 A36-H-B12A37-H-B12 A38-H-B12 A39-H-B12 A40-H-B12 A41-H-B12 A42-H-B12 A36-H-B13A37-H-B13 A38-H-B13 A39-H-B13 A40-H-B13 A41-H-B13 A42-H-B13 A36-H-B14A37-H-B14 A38-H-B14 A39-H-B14 A40-H-B14 A41-H-B14 A42-H-B14 A36-H-B15A37-H-B15 A38-H-B15 A39-H-B15 A40-H-B15 A41-H-B15 A42-H-B15 A43-H-B1A44-H-B1 A45-H-B1 A46-H-B1 A47-H-B1 A48-H-B1 A49-H-B1 A43-H-B2 A44-H-B2A45-H-B2 A46-H-B2 A47-H-B2 A48-H-B2 A49-H-B2 A43-H-B3 A44-H-B3 A45-H-B3A46-H-B3 A47-H-B3 A48-H-B3 A49-H-B3 A43-H-B4 A44-H-B4 A45-H-B4 A46-H-B4A47-H-B4 A48-H-B4 A49-H-B4 A43-H-B5 A44-H-B5 A45-H-B5 A46-H-B5 A47-H-B5A48-H-B5 A49-H-B5 A43-H-B6 A44-H-B6 A45-H-B6 A46-H-B6 A47-H-B6 A48-H-B6A49-H-B6 A43-H-B7 A44-H-B7 A45-H-B7 A46-H-B7 A47-H-B7 A48-H-B7 A49-H-B7A43-H-B8 A44-H-B8 A45-H-B8 A46-H-B8 A47-H-B8 A48-H-B8 A49-H-B8 A43-H-B9A44-H-B9 A45-H-B9 A46-H-B9 A47-H-B9 A48-H-B9 A49-H-B9 A43-H-B10A44-H-B10 A45-H-B10 A46-H-B10 A47-H-B10 A48-H-B10 A49-H-B10 A43-H-B11A44-H-B11 A45-H-B11 A46-H-B11 A47-H-B11 A48-H-B11 A49-H-B11 A43-H-B12A44-H-B12 A45-H-B12 A46-H-B12 A47-H-B12 A48-H-B12 A49-H-B12 A43-H-B13A44-H-B13 A45-H-B13 A46-H-B13 A47-H-B13 A48-H-B13 A49-H-B13 A43-H-B14A44-H-B14 A45-H-B14 A46-H-B14 A47-H-B14 A48-H-B14 A49-H-B14 A43-H-B15A44-H-B15 A45-H-B15 A46-H-B15 A47-H-B15 A48-H-B15 A49-H-B15 A50-H-B1A51-H-B1 A52-H-B1 A53-H-B1 A54-H-B1 A55-H-B1 A56-H-B1 A50-H-B2 A51-H-B2A52-H-B2 A53-H-B2 A54-H-B2 A55-H-B2 A56-H-B2 A50-H-B3 A51-H-B3 A52-H-B3A53-H-B3 A54-H-B3 A55-H-B3 A56-H-B3 A50-H-B4 A51-H-B4 A52-H-B4 A53-H-B4A54-H-B4 A55-H-B4 A56-H-B4 A50-H-B5 A51-H-B5 A52-H-B5 A53-H-B5 A54-H-B5A55-H-B5 A56-H-B5 A50-H-B6 A51-H-B6 A52-H-B6 A53-H-B6 A54-H-B6 A55-H-B6A56-H-B6 A50-H-B7 A51-H-B7 A52-H-B7 A53-H-B7 A54-H-B7 A55-H-B7 A56-H-B7A50-H-B8 A51-H-B8 A52-H-B8 A53-H-B8 A54-H-B8 A55-H-B8 A56-H-B8 A50-H-B9A51-H-B9 A52-H-B9 A53-H-B9 A54-H-B9 A55-H-B9 A56-H-B9 A50-H-B10A51-H-B10 A52-H-B10 A53-H-B10 A54-H-B10 A55-H-B10 A56-H-B10 A50-H-B11A51-H-B11 A52-H-B11 A53-H-B11 A54-H-B11 A55-H-B11 A56-H-B11 A50-H-B12A51-H-B12 A52-H-B12 A53-H-B12 A54-H-B12 A55-H-B12 A56-H-B12 A50-H-B13A51-H-B13 A52-H-B13 A53-H-B13 A54-H-B13 A55-H-B13 A56-H-B13 A50-H-B14A51-H-B14 A52-H-B14 A53-H-B14 A54-H-B14 A55-H-B14 A56-H-B14 A50-H-B15A51-H-B15 A52-H-B15 A53-H-B15 A54-H-B15 A55-H-B15 A56-H-B15 A57-H-B1A58-H-B1 A59-H-B1 A60-H-B1 A61-H-B1 A62-H-B1 A63-H-B1 A57-H-B2 A58-H-B2A59-H-B2 A60-H-B2 A61-H-B2 A62-H-B2 A63-H-B2 A57-H-B3 A58-H-B3 A59-H-B3A60-H-B3 A61-H-B3 A62-H-B3 A63-H-B3 A57-H-B4 A58-H-B4 A59-H-B4 A60-H-B4A61-H-B4 A62-H-B4 A63-H-B4 A57-H-B5 A58-H-B5 A59-H-B5 A60-H-B5 A61-H-B5A62-H-B5 A63-H-B5 A57-H-B6 A58-H-B6 A59-H-B6 A60-H-B6 A61-H-B6 A62-H-B6A63-H-B6 A57-H-B7 A58-H-B7 A59-H-B7 A60-H-B7 A61-H-B7 A62-H-B7 A63-H-B7A57-H-B8 A58-H-B8 A59-H-B8 A60-H-B8 A61-H-B8 A62-H-B8 A63-H-B8 A57-H-B9A58-H-B9 A59-H-B9 A60-H-B9 A61-H-B9 A62-H-B9 A63-H-B9 A57-H-B10A58-H-B10 A59-H-B10 A60-H-B10 A61-H-B10 A62-H-B10 A63-H-B10 A57-H-B11A58-H-B11 A59-H-B11 A60-H-B11 A61-H-B11 A62-H-B11 A63-H-B11 A57-H-B12A58-H-B12 A59-H-B12 A60-H-B12 A61-H-B12 A62-H-B12 A63-H-B12 A57-H-B13A58-H-B13 A59-H-B13 A60-H-B13 A61-H-B13 A62-H-B13 A63-H-B13 A57-H-B14A58-H-B14 A59-H-B14 A60-H-B14 A61-H-B14 A62-H-B14 A63-H-B14 A57-H-B15A58-H-B15 A59-H-B15 A60-H-B15 A61-H-B15 A62-H-B15 A63-H-B15 A64-H-B1A65-H-B1 A66-H-B1 A67-H-B1 A68-H-B1 A69-H-B1 A70-H-B1 A64-H-B2 A65-H-B2A66-H-B2 A67-H-B2 A68-H-B2 A69-H-B2 A70-H-B2 A64-H-B3 A65-H-B3 A66-H-B3A67-H-B3 A68-H-B3 A69-H-B3 A70-H-B3 A64-H-B4 A65-H-B4 A66-H-B4 A67-H-B4A68-H-B4 A69-H-B4 A70-H-B4 A64-H-B5 A65-H-B5 A66-H-B5 A67-H-B5 A68-H-B5A69-H-B5 A70-H-B5 A64-H-B6 A65-H-B6 A66-H-B6 A67-H-B6 A68-H-B6 A69-H-B6A70-H-B6 A64-H-B7 A65-H-B7 A66-H-B7 A67-H-B7 A68-H-B7 A69-H-B7 A70-H-B7A64-H-B8 A65-H-B8 A66-H-B8 A67-H-B8 A68-H-B8 A69-H-B8 A70-H-B8 A64-H-B9A65-H-B9 A66-H-B9 A67-H-B9 A68-H-B9 A69-H-B9 A70-H-B9 A64-H-B10A65-H-B10 A66-H-B10 A67-H-B10 A68-H-B10 A69-H-B10 A70-H-B10 A64-H-B11A65-H-B11 A66-H-B11 A67-H-B11 A68-H-B11 A69-H-B11 A70-H-B11 A64-H-B12A65-H-B12 A66-H-B12 A67-H-B12 A68-H-B12 A69-H-B12 A70-H-B12 A64-H-B13A65-H-B13 A66-H-B13 A67-H-B13 A68-H-B13 A69-H-B13 A70-H-B13 A64-H-B14A65-H-B14 A66-H-B14 A67-H-B14 A68-H-B14 A69-H-B14 A70-H-B14 A64-H-B15A65-H-B15 A66-H-B15 A67-H-B15 A68-H-B15 A69-H-B15 A70-H-B15

EXAMPLE 13-(5,6-Dimethyl-1H-benzoimidazol-2-yl)-2,4,5,7-tetrahydro-pyrazolo[3,4-c]pyridine-6-carboxylicacid 2-phenylethylamide

3-(5,6-Dimethyl-1H-benzoimidazol-2-yl)-2,4,5,7-tetrahydropyrazolo-[3,4-c]pyridine-6-carboxylicacid 2-phenylethylamide can be prepared in the following way:

10 mg of3-(5,6-dimethyl-1H-benzimidazol-2-yl)-4,5,6,7-tetrahydro-2H-pyrazolo[3,4-c]pyridineare suspended in 0.3 ml of tetrahydrofuran. 7.8 μl of 2-phenylethylisocyanate are added and the reaction mixture is stirred at ambienttemperature for 20 hours and then concentrated under reduced pressure.

The evaporation residue is purified by LC/MS (method B). Afterpurification by LC/MS, the fractions containing the3-(5,6-dimethyl-1H-benzoimidazol-2-yl)-2,4,5,7-tetrahydropyrazolo[3,4-c]pyridine-6-carboxylicacid 2-phenyl-ethylamide are combined and loaded onto SCX phase (500 mgof CUBCX1-HL phase). The SCX phase is subsequently washed with methanoland then extracted with a solution of 2M ammonia in methanol. Theextraction solution obtained is then concentrated under reducedpressure. 1.2 mg of3-(5,6-dimethyl-1H-benzoimidazol-2-yl)-2,4,5,7-tetrahydropyrazolo[3,4-c]-pyridine-6-carboxylicacid 2-phenylethylamide are thus obtained in the form of a white powder,the characteristics of which are as follows:

LC/MS (method A): molecular ion detected: 415.29; retention time=3.48minutes

EXAMPLE 23-(5,6-Dimethyl-1H-benzoimidazol-2-yl)-6-methanesulphonyl-4,5,6,7-tetrahydro-2H-pyrazolo[3,4-c]pyridine

3-(5,6-Dimethyl-1H-benzoimidazol-2-yl)-6-methanesulphonyl-4,5,6,7-tetrahydro-2H-pyrazolo[3,4-c]pyridinecan be prepared in the following way:

10 mg of3-(5,6-dimethyl-1H-benzimidazol-2-yl)-4,5,6,7-tetrahydro-2H-pyrazolo[3,4-c]pyridineare suspended in 0.3 ml of dichloromethane. 15.8 μl of triethylamine areadded, along with 4.5 μl of methanesulphonyl chloride. The reactionmixture is stirred at ambient temperature for 20 hours and is thenconcentrated under reduced pressure.

The evaporation residue is purified by LC/MS (method B). Afterpurification by LC/MS, the fractions containing3-(5,6-dimethyl-1H-benzoimidazol-2-yl)-6-methanesulphonyl-4,5,6,7-tetrahydro-2H-pyrazolo[3,4-c]pyridineare combined and loaded onto SCX phase (500 mg of CUBCX1-HL phase). TheSCX phase is subsequently washed with methanol and then extracted with asolution of 2M ammonia in methanol. The extraction solution obtained isthen concentrated under reduced pressure. 4.2 mg of3-(5,6-dimethyl-1H-benzoimidazol-2-yl)-6-methanesulphonyl-4,5,6,7-tetrahydro-2H-pyrazolo-[3,4-c]pyridineare thus obtained in the form of a white powder, the characteristics ofwhich are as follows:

LC/MS (method A): molecular ion detected: 346.30; retention time=3.08minutes.

¹H NMR (300 MHz, (CD₃)₂SO, δ in ppm): 2.32 (broad s: 6H); 3.02 (s: 3H);3.03 (mt: 2H); 3.52 (broad t, J=5 Hz: 2H); 4.45 (broad s: 2H); 7.24(broad s: 1H); 7.42 (broad s: 1H); 12.45 (unresolved peak: 1H); 13.07(unresolved peak: 1H).

EXAMPLE 3[3-(5,6-Dimethyl-1H-benzoimidazol-2-yl)-2,4,5,7-tetrahydro-pyrazolo[3,4-c]pyridin-6-yl]-3-pyridinylmethanone

[3-(5,6-Dimethyl-1H-benzoimidazol-2-yl)-2,4,5,7-tetrahydropyrazolo-[3,4-c]pyridin-6-yl]-3-pyridinylmethanonecan be prepared in the following way:

10 mg of3-(5,6-dimethyl-1H-benzimidazol-2-yl)-4,5,6,7-tetrahydro-2H-pyrazolo[3,4-c]pyridineare suspended in 0.3 ml of DMF. 6.9 mg of nicotinic acid are added,followed by 7.6 mg of HOBT and 8.7 μl of diisopropylcarbo-diimide.

The reaction mixture is stirred at ambient temperature for 20 hours andis then concentrated under reduced pressure.

The evaporation residue is purified by LC/MS (method B). Afterpurification by LC/MS, the fractions containing the[3-(5,6-dimethyl-1H-benzoimidazol-2-yl)-2,4,5,7-tetrahydropyrazolo[3,4-c]pyridin-6-yl]-3-pyridinylmethanoneare combined and loaded onto SCX phase (500 mg of CUBCX1-HL phase). TheSCX phase is subsequently washed with methanol and then extracted with asolution of 2M ammonia in methanol. The extraction solution obtained isthen concentrated under reduced pressure. 5.3 mg of[3-(5,6-dimethyl-1H-benzoimidazol-2-yl)-2,4,5,7-tetrahydropyrazolo[3,4-c]pyridin-6-yl]-3-pyridinyl-methanoneare thus obtained in the form of a white powder, the characteristics ofwhich are as follows:

LC/MS (method A): molecular ion detected: 373.31; retention time=2.87minutes

¹H NMR (400 MHz, (CD₃)₂SO, at a temperature of 373K, δ in ppm): 2.35 (s:6H); from 2.90 to 3.10 (mt: 2H); 3.76 (unresolved peak: 2H); 4.76 (broads: 2H); 7.27 (unresolved peak: 1H); 7.40 (unresolved peak: 1H); 7.51(dd, J=8 and 5 Hz: 1H); 7.90 (broad d, J=8 Hz: 1H); 8.70 (mt: 2H); from11.80 to 12.20 (broad unresolved peak: 1H); from 12.50 to 13.00 (broadunresolved peak: 1H).

EXAMPLE 46-(3-Chlorobenzyl)-3-(5,6-dimethyl-1H-benzoimidazol-2-yl)-4,5,6,7-tetrahydro-2H-pyrazolo[3,4-c]pyridine

6-(3-Chlorobenzyl)-3-(5,6-dimethyl-1H-benzoimidazol-2-yl)4,5,6,7-tetrahydro-2H-pyrazolo[3,4-c]pyridinecan be prepared in the following way:

10 mg of3-(5,6-dimethyl-1H-benzimidazol-2-yl)-4,5,6,7-tetrahydro-2H-pyrazolo[3,4-c]pyridineare suspended in 0.3 ml of methanol. 12.7 μl of 3-chlorobenzaldehyde areadded, followed by 4.7 mg of NaBH₃CN. The reaction mixture is stirred atambient temperature for 20 hours and is then concentrated under reducedpressure.

The evaporation residue is purified by LC/MS (method B). Afterpurification by LC/MS, the fractions containing the6-(3-chlorobenzyl)-3-(5,6-dimethyl-1H-benzoimidazol-2-yl)-4,5,6,7-tetrahydro-2H-pyrazolo[3,4-c]pyridineare combined and loaded onto a SCX phase (500 mg of CUBCX1-HL phase).The SCX phase is subsequently washed with methanol and then extractedwith a solution of 2M ammonia in methanol. The extraction solutionobtained is then concentrated under reduced pressure. 4 mg of6-(3-chlorobenzyl)-3-(5,6-dimethyl-1H-benzoimidazol-2-yl)4,5,6,7-tetrahydro-2H-pyrazolo-[3,4-c]pyridineare thus obtained in the form of a white powder, the characteristics ofwhich are as follows:

LC/MS (method A): molecular ion detected: 392.26; retention time 3.18minutes.

¹H NMR (400 MHz, (CD₃)₂SO, at a temperature of 373K, δ in ppm): 2.35 and2.36 (2 s: 6H in total); 2.83 (t, J=5.5 Hz: 2H); from 2.90 to 3.00 (mt:2H); 3.62 (broad s: 2H); 3.78 (s: 2H); from 7.25 to 7.50 (mt: 6H); 11.91(unresolved peak: 1H); from 12.30 to 12.60 (broad unresolved peak: 1H).

EXAMPLE 5[3-(1H-Benzoimidazol-2-yl)-2,4,5,7-tetrahydropyrazolo[3,4-c]-pyridin-6-yl]-3-pyridinylmethanone

[3-(1H-Benzoimidazol-2-yl)-2,4,5,7-tetrahydropyrazolo[3,4-c]pyridin-6-yl]-3-pyridinylmethanonecan be prepared in the following way:

15 mg of3-(1H-benzimidazol-2-yl)-4,5,6,7-tetrahydro-2H-pyrazolo[3,4-c]-pyridinehydrochloride are suspended in 0.5 ml of DMF. 24.3 mg ofdiisopropylethylamine are added, followed by 12.7 mg of HOBT, 11.9 mg ofdiisopropylcarbodiimide and 11.6 mg of nicotinic acid. The reactionmixture is stirred at ambient temperature for 20 hours and thenconcentrated under reduced pressure.

The evaporation residue is purified by LC/MS (method B). Afterpurification by LC/MS, the fractions containing the[3-(1H-benzoimidazol-2-yl)-2,4,5,7-tetrahydropyrazolo[3,4-c]pyridin-6-yl]-3-pyridinylmethanoneare combined and loaded onto SCX phase (500 mg of CUBCX1-HL phase). TheSCX phase is subsequently washed with methanol and then extracted with asolution of 2M ammonia in methanol. The extraction solution obtained isthen concentrated under reduced pressure. 7.7 mg of[3-(1H-benzoimidazol-2-yl)-2,4,5,7-tetrahydropyrazolo[3,4-c]pyridin-6-yl]-3-pyridinylmethanoneare thus obtained in the form of a white powder, the characteristics ofwhich are as follows:

LC/MS (method A): molecular ion detected: 345.22; retention time=1.95minutes

EXAMPLE 66-(3-Chlorobenzyl)-3-(1H-benzoimidazol-2-yl)-4,5,6,7-tetrahydro-2H-pyrazolo[3,4-c]pyridine

6-(3-Chlorobenzyl)-3-(1H-benzoimidazol-2-yl)-4,5,6,7-tetrahydro-2H-pyrazolo[3,4-c]pyridinecan be prepared in the following way:

15 mg of3-(1H-benzimidazol-2-yl)-4,5,6,7-tetrahydro-2H-pyrazolo[3,4-c]-pyridinehydrochloride are suspended in 0.5 ml of methanol. 26.5 mg of3-chlorobenzaldehyde are added, followed by 7.9 mg of NaBH₃CN. Thereaction mixture is stirred at ambient temperature for 20 hours and isthen concentrated under reduced pressure.

The evaporation residue is purified by LC/MS (method B). Afterpurification by LC/MS, the fractions containing the6-(3-chlorobenzyl)-3-(1H-benzoimidazol-2-yl)-4,5,6,7-tetrahydro-2H-pyrazolo[3,4-c]pyridineare combined and loaded onto SCX phase (500 mg of CUBCX1-HL phase). TheSCX phase is subsequently washed with methanol and extracted with asolution of 2M ammonia in methanol. The extraction solution obtained isthen concentrated under reduced pressure. 6.9 mg of6-(3-chlorobenzyl)-3-(1H-benzoimidazol-2-yl)-4,5,6,7-tetrahydro-2H-pyrazolo[3,4-c]pyridineare thus obtained in the form of a white powder, the characteristics ofwhich are as follows:

LC/MS (method A): molecular ion detected: 364.22; retention time=2.19minutes

EXAMPLE 7 Preparation of an Amide Library

The amide library can be prepared in the following way:

The 19 acids (Table 4) are weighed and placed in 19 individual testtubes.

TABLE 4 Acids used Entry Name Amount 1 ISOBUTYRIC ACID 3.3 mg 2 BENZOICACID 4.6 mg 3 2,3-DICHLOROBENZOIC ACID 7.1 mg 4 PHENYLACETIC ACID 5.1 mg5 ACETIC ACID 2.2 mg 6 CYCLOPROPANECARBOXYLIC ACID 3.2 mg 72-CHLOROBENZOIC ACID 5.9 mg 8 3-CHLOROBENZOIC ACID 5.9 mg 94-CHLOROBENZOIC ACID 5.9 mg 10 ISOVALERIC ACID 3.8 mg 11 HYDROCINNAMICACID 5.6 mg 12 VINYLACETIC ACID 3.2 mg 13 BUTYRIC ACID 3.3 mg 142-FUROIC ACID 4.2 mg 15 PIVALIC ACID 3.8 mg 16 N,N-DIMETHYLGLYCINE 3.9mg 17 VALERIC ACID 3.8 mg 18 THIOPHENE-2-CARBOXYLIC ACID 4.8 mg 194-METHYLSULPHONYLBENZOIC ACID 7.5 mg

152 mg of HOBT and 142 mg of diisopropylcarbodiimide are solubilized in12 ml of DMF and the solution obtained is distributed in each of the 19test tubes, at a rate of 600 μl per tube.

200 mg of3-(5,6-dimethyl-1H-benzimidazol-2-yl)-4,5,6,7-tetrahydro-2H-pyrazolo[3,4-c]pyridinehydrochloride are suspended in 4 ml of DMF in the presence of 290 mg ofN,N-diisopropylethylamine, and the suspension obtained is distributedinto each of the 19 test tubes, at a rate of 200 μl per tube.

The 19 reaction mixtures are shaken by means of orbital shaking atambient temperature for 20 hours.

For each reaction mixture, a 10 μl sample is taken and diluted in 40 μlof DMSO (Gilson Liquid Handler Quad-Z 215). Each sample in solution inDMSO thus obtained is analysed by LC/MS (method A).

The 19 reaction mixtures are then evaporated to dryness and theevaporation residues are each solubilized in 500 μl of DMSO, and thesolutions obtained are then purified by LC/MS (method B).

After purification by LC/MS, the fractions containing the desiredcompounds are (optionally combined) loaded onto SCX phase (500 mg ofCUBCX1-HL phase). The SCX phases are subsequently washed with methanoland then extracted with a solution of 2M ammonia in methanol. Theextraction solutions are collected in tared glass tubes, evaporated todryness (Savant AES 2000 or Genevac HT8 centrifugal evaporator), weighed(Mettler Toledo Automated Workstation LA200) and diluted to 10 mM inDMSO (Gilson Liquid Handler Quad-Z 215). Each solution obtained isanalysed by LC/MS (method A).

The following compounds (Table 5) were isolated and characterized bymeans of their retention time and molecular peak in mass spectrometry(method A).

TABLE 5 Amide library obtained Amount of Retention Molecular producttime ion Entry Name obtained (minutes) detected 11-[3-(5,6-Dimethyl-1H-benzoimidazol- 5.8 mg 3.08 338.232-yl)-2,4,5,7-tetrahydropyrazolo- [3,4-c]pyridin-6-yl]-2-methylpropan-1-one 2 [3-(5,6-Dimethyl-1H-benzoimidazol-2- 6.8 mg 2.68 372.21yl)-2,4,5,7-tetrahydropyrazolo[3,4-c]- pyridin-6-yl]phenylmethanone 3(2,3-Dichlorophenyl)-[3-(5,6-dimethyl- 12 mg 3.05 440.131H-benzoimidazol-2-yl)-2,4,5,7- tetrahydropyrazolo[3,4-c]pyridin-6-yl]-methanone 4 1-[3-(5,6-Dimethyl-1H-benzoimidazol- 7.9 mg 2.99 386.232-yl)-2,4,5,7-tetrahydropyrazolo- [3,4-c]pyridin-6-yl]-2-phenylethanone5 1-[3-(5,6-Dimethyl-1H-benzoimidazol- 2.7 mg 2.4 310.192-yl)-2,4,5,7-tetrahydropyrazolo- [3,4-c]pyridin-6-yl]ethanone 6Cyclopropyl-[3-(5,6-dimethyl-1H- 3.4 mg 2.57 336.21benzoimidazol-2-yl)-2,4,5,7- tetrahydropyrazolo[3,4-c]pyridin-6-yl]-methanone 7 (2-Chlorophenyl)-[3-(5,6-dimethyl-1H- 11.2 mg 2.97 406.18benzoimidazol-2-yl)-2,4,5,7- tetrahydropyrazolo[3,4-c]pyridin-6-yl]-methanone 8 (3-Chlorophenyl)-[3-(5,6-dimethyl-1H- 12.1 mg 3.31 406.16benzoimidazol-2-yl)-2,4,5,7- tetrahydropyrazolo[3,4-c]pyridin-6-yl]-methanone 9 (4-Chlorophenyl)-[3-(5,6-dimethyl-1H- 11.5 mg 3.51 406.17benzoimidazol-2-yl)-2,4,5,7- tetrahydropyrazolo[3,4-c]pyridin-6-yl]-methanone 10 1-[3-(5,6-Dimethyl-1H-benzoimidazol- 4.8 mg 2.72 352.242-yl)-2,4,5,7-tetrahydropyrazolo[3,4-c]-pyridin-6-yl]-3-methylbutan-1-one 111-[3-(5,6-Dimethyl-1H-benzoimidazol- 11.9 mg 2.95 400.242-yl)-2,4,5,7-tetrahydropyrazolo[3,4-c]-pyridin-6-yl]-3-phenylpropan-1-one 121-[3-(5,6-Dimethyl-1H-benzoimidazol- 10.1 mg 2.72 336.222-yl)-2,4,5,7-tetrahydropyrazolo- [3,4-c]pyridin-6-yl]but-3-en-1-one 131-[3-(5,6-Dimethyl-1H-benzoimidazol- 7 mg 2.66 338.232-yl)-2,4,5,7-tetrahydropyrazolo- [3,4-c]pyridin-6-yl]butan-1-one 14[3-(5,6-Dimethyl-1H-benzoimidazol-2- 9.5 mg 2.67 362.19yl)-2,4,5,7-tetrahydropyrazolo[3,4-c]- pyridin-6-yl]furan-2-yl-methanone15 1-[3-(5,6-Dimethyl-1H-benzoimidazol- 9.3 mg 2.8 352.242-yl)-2,4,5,7-tetrahydropyrazolo[3,4-c]-pyridin-6-yl]-2,2-dimethylpropan-1-one 162-Dimethylamino-1-[3-(5,6-dimethyl- 4.7 mg 2.55 353.231H-benzoimidazol-2-yl)-2,4,5,7- tetrahydro-pyrazolo[3,4-c]pyridin-6-yl]ethanone 17 1-[3-(5,6-Dimethyl-1H-benzoimidazol- 5.4 mg 2.78 352.242-yl)-2,4,5,7-tetrahydropyrazolo[3,4-c]- pyridin-6-yl]pentan-1-one 18[3-(5,6-Dimethyl-1H-benzoimidazol-2- 7.2 mg 2.75 378.17yl)-2,4,5,7-tetrahydropyrazolo[3,4-c]-pyridin-6-yl]thiophen-2-yl-methanone 19[3-(5,6-Dimethyl-1H-benzoimidazol-2- 14.3 mg 2.79 450.19yl)-2,4,5,7-tetrahydropyrazolo[3,4-c]-pyridin-6-yl]-(4-methanesulphonyl- phenyl)methanone

EXAMPLE 8 Preparation of a Sulphonamide Library

The sulphonamide library can be prepared in the following way:

190 mg of3-(5,6-dimethyl-1H-benzimidazol-2-yl)-4,5,6,7-tetrahydro-2H-pyrazolo[3,4-c]pyridinehydrochloride are suspended in 2 ml of dichloromethane in the presenceof 150 μl of triethylamine, and the suspension obtained is distributedinto 17 test tubes, at a rate of 500 μl per tube. The 17 sulphonylchlorides (Table 6) are weighed and added to each of the 17 test tubes.

TABLE 6 Sulphonyl chlorides used Entry Name Quantity 1 BENZENESULPHONYLCHLORIDE 9.9 mg 2 ALPHA-TOLUENESULPHONYL 10.7 mg CHLORIDE 32,3-DICHLOROBENZENE- 13.8 mg SULPHONYL CHLORIDE 44-CHLOROBENZENESULPHONYL 11.9 mg CHLORIDE 5 2,2,2-TRIFLUOROETHANE- 10.2mg SULPHONYL CHLORIDE 6 ETHANESULPHONYL CHLORIDE 7.2 mg 71-PROPANESULPHONYL CHLORIDE 8 mg 8 1-BUTANESULPHONYL CHLORIDE 8.8 mg 92-CHLOROBENZENESULPHONYL 11.9 mg CHLORIDE 10 3-CHLOROBENZENESULPHONYL11.9 mg CHLORIDE 11 [(4-FLUOROPHENYL)METHYL]- 11.7 mg SULPHONYL CHLORIDE12 4-METHOXYBENZENESULPHONYL 11.6 mg CHLORIDE 13 P-TOLUENESULPHONYLCHLORIDE 10.7 mg 14 O-TOLUENESULPHONYL CHLORIDE 10.7 mg 153-METHYLBENZENESULPHONYL 10.7 mg CHLORIDE 16 3-METHOXYBENZENESULPHONYL11.6 mg CHLORIDE 17 2-METHOXY-4-METHYLBENZENE- 12.4 mg SULPHONYLCHLORIDE

The 17 reaction mixtures are shaken by means of orbital shaking atambient temperature for 20 h.

For each reaction mixture, a 10 μl sample is taken and diluted in 40 μlof DMSO (Gilson Liquid Handler Quad-Z 215). Each sample in solution inDMSO thus obtained is analysed by LC/MS (method A).

The 17 reaction mixtures are then evaporated to dryness and theevaporation residues are each solubilized in 1 ml of DMSO in thepresence of a drop of an aqueous 5N hydrochloric acid solution, and thesolutions obtained are purified by LC/MS (method B). After purificationby LC/MS, the fractions containing the desired compounds are (optionallycombined) loaded onto SCX phase (500 mg of CUBCX1-HL phase). The SCXphases are subsequently washed with methanol and then extracted with asolution of 2M ammonia in methanol. The extraction solutions arecollected in tared glass tubes, evaporated to dryness (Savant AES 2000or Genevac HT8 centrifugal evaporator), weighed (Mettler ToledoAutomated Workstation LA200) and diluted to 10 mM in DMSO (Gilson LiquidHandler Quad-Z 215). Each solution obtained is analysed by LC/MS (methodA).

The following compounds (Table 7) were isolated and characterized bymeans of their retention time and molecular peak in mass spectrometry(method A).

TABLE 7 Sulphonamide library obtained Amount Retention Molecular ofproduct time ion Entry Name obtained (minutes) detected 16-Benzenesulphonyl-3-(5,6-dimethyl- 1.4 mg 3.41 408.181H-benzoimidazol-2-yl)-4,5,6,7- tetrahydro-2H-pyrazolo[3,4-c]pyridine 23-(5,6-Dimethyl-1H-benzoimidazol-2-yl)- 0.7 mg 3.51 422.26-phenylmethanesulphonyl-4,5,6,7- tetrahydro-2H-pyrazolo[3,4-c]pyridine3 6-(2,3-Dichlorobenzenesulphonyl)-3- 6.4 mg 3.25 476.1(5,6-dimethyl-1H-benzoimidazol-2-yl)- 4,5,6,7-tetrahydro-2H-pyrazolo-[3,4-c]pyridine 4 6-(4-Chlorobenzenesulphonyl)-3-(5,6- 5.9 mg 3.19442.12 dimethyl-1H-benzoimidazol-2-yl)- 4,5,6,7-tetrahydro-2H-pyrazolo-[3,4-c]pyridine 5 3-(5,6-Dimethyl-1H-benzoimidazol-2-yl)- 1.7 mg 3.06414.14 6-(2,2,2-trifluoroethanesulphonyl)-4,5,6,7-tetrahydro-2H-pyrazolo- [3,4-c]pyridine 63-(5,6-Dimethyl-1H-benzoimidazol-2-yl)- 5.2 mg 2.63 360.176-ethanesulphonyl-4,5,6,7-tetrahydro- 2H-pyrazolo[3,4-c]pyridine 73-(5,6-Dimethyl-1H-benzoimidazol-2-yl)- 4.3 mg 2.8 374.196-(propane-1-sulphonyl)-4,5,6,7- tetrahydro-2H-pyrazolo[3,4-c]pyridine 86-(Butane-1-sulphonyl)-3-(5,6-dimethyl- 5.6 mg 2.94 388.21H-benzoimidazol-2-yl)-4,5,6,7- tetrahydro-2H-pyrazolo[3,4-c]pyridine 96-(2-Chlorobenzenesulphonyl)-3-(5,6- 5.6 mg 3.38 442.13dimethyl-1H-benzoimidazol-2-yl)- 4,5,6,7-tetrahydro-2H-pyrazolo[3,4-c]-pyridine 10 6-(3-Chlorobenzenesulphonyl)-3-(5,6- 6.9 mg 3.71 442.13dimethyl-1H-benzoimidazol-2-yl)- 4,5,6,7-tetrahydro-2H-pyrazolo[3,4-c]-pyridine 11 3-(5,6-Dimethyl-1H-benzoimidazol-2-yl)- 0.7 mg 3.05 440.186-(4-fluorophenylmethanesulphonyl)-4,5,6,7-tetrahydro-2H-pyrazolo[3,4-c]- pyridine 123-(5,6-Dimethyl-1H-benzoimidazol-2-yl)- 7.5 mg 2.99 438.196-(4-methoxybenzenesulphonyl)- 4,5,6,7-tetrahydro-2H-pyrazolo[3,4-c]-pyridine 13 3-(5,6-Dimethyl-1H-benzoimidazol-2-yl)- 7.3 mg 3.22 422.26-(toluene-4-sulphonyl)-4,5,6,7- tetrahydro-2H-pyrazolo[3,4-c]pyridine14 3-(5,6-Dimethyl-1H-benzoimidazol-2-yl)- 4.8 mg 3.16 422.196-(toluene-2-sulphonyl)-4,5,6,7- tetrahydro-2H-pyrazolo[3,4-c]pyridine15 3-(5,6-Dimethyl-1H-benzoimidazol-2-yl)- 5.1 mg 3.13 422.196-(toluene-3-sulphonyl)-4,5,6,7- tetrahydro-2H-pyrazolo[3,4-c]pyridine16 3-(5,6-Dimethyl-1H-benzoimidazol-2-yl)- 6.9 mg 3.07 438.186-(3-methoxybenzenesulphonyl)- 4,5,6,7-tetrahydro-2H-pyrazolo[3,4-c]-pyridine 17 3-(5,6-Dimethyl-1H-benzoimidazol-2-yl)- 0.8 mg 3.34 452.196-(2-methoxy-4-methyl-benzene- sulphonyl)-4,5,6,7-tetrahydro-2H-pyrazolo[3,4-c]pyridine

EXAMPLE 9 Preparation of an Amine Library

The amine library can be prepared in the following way:

180 mg of3-(5,6-dimethyl-1H-benzimidazol-2-yl)-4,5,6,7-tetrahydro-2H-pyrazolo[3,4-c]pyridinehydrochloride are suspended in 2.7 ml of methanol and the suspensionobtained is distributed into 16 test tubes, at a rate of 150 μl pertube.

The 16 aldehyde (Table 8) are weighed and added to each of the 16 testtubes.

TABLE 8 Aldehydes used Entry Name Quantity 1 ISOBUTYRALDEHYDE 8.1 mg 2FORMALDEHYDE 3.4 mg 3 BENZALDEHYDE 11.9 mg 4 PHENYLACETALDEHYDE 13.5 mg5 2,3-DICHLOROBENZALDEHYDE 19.6 mg 6 FURFURAL 10.8 mg 74-CHLOROBENZALDEHYDE 15.8 mg 8 2-THIOPHENECARBOXALDEHYDE 12.6 mg 9NICOTINALDEHYDE 12 mg 10 TRIMETHYLACETALDEHYDE 9.7 mg 11 ACETALDEHYDE4.9 mg 12 ISOVALERALDEHYDE 9.7 mg 13 PROPIONALDEHYDE 6.5 mg 143-PHENYLPROPIONALDEHYDE 15.1 mg 15 BUTYRALDEHYDE 8.1 mg 16CYCLOPROPANECARBOXALDEHYDE 7.9 mg

A solution of 85 mg of NaBH₃CN in 2.7 ml of methanol is then alsodistributed into the 16 test tubes, at a rate of 150 μl per tube. The 16reaction mixtures are shaken by means of orbital shaking at ambienttemperature for 20 h. 100 μl of methanol are then added to each of the16 tubes.

For each reaction mixture, a 10 μl sample is taken and diluted in 40 μlof DMSO (Gilson Liquid Handler Quad-Z 215). Each sample in solution inDMSO thus obtained is analysed by LC/MS (method A).

The 16 reaction mixtures are then evaporated to dryness and theevaporation residues are each solubilized in 500 μl of DMSO and filteredthrough sintered glass, and the residual solutions are then purified byLC/MS (method B). After purification by LC/MS, the fractions containingthe desired compounds are (optionally combined) loaded onto SCX phase(500 mg of CUBCX1-HL phase). The SCX phases are subsequently washed withmethanol and then extracted with a solution of 2M ammonia in methanol.The extraction solutions are collected in tared glass tubes, evaporatedto dryness (Savant AES 2000 or Genevac HT8 centrifugal evaporator),weighed (Mettler Toledo Automated Workstation LA200) and diluted to 10mM in DMSO (Gilson Liquid Handler Quad-Z 215). Each solution obtained isanalysed by LC/MS (method A).

The following compounds (Table 9) were isolated and characterized bymeans of their retention time and molecular peak in mass spectrometry(method A).

TABLE 9 Amine library obtained Amount of Retention Molecular producttime ion Entry Name obtained (minutes) detected 13-(5,6-Dimethyl-1H-benzoimidazol-2- 5.9 mg 2.62 324.32yl)-6-isobutyl-4,5,6,7-tetrahydro-2H- pyrazolo[3,4-c]pyridine 23-(5,6-Dimethyl-1H-benzoimidazol-2- 3.5 mg 2.49 282.29yl)-6-methyl-4,5,6,7-tetrahydro-2H- pyrazolo[3,4-c]pyridine 36-Benzyl-3-(5,6-dimethyl-1H-benzo- 8.2 mg 2.74 358.3imidazol-2-yl)-4,5,6,7-tetrahydro-2H- pyrazolo[3,4-c]pyridine 43-(5,6-Dimethyl-1H-benzoimidazol-2- 6.4 mg 2.84 372.32yl)-6-phenethyl-4,5,6,7-tetrahydro-2H- pyrazolo[3,4-c]pyridine 56-(2,3-Dichlorobenzyl)-3-(5,6-dimethyl- 8.6 mg 2.95 426.231H-benzoimidazol-2-yl)-4,5,6,7- tetrahydro-2H-pyrazolo[3,4-c]pyridine 63-(5,6-Dimethyl-1H-benzoimidazol-2- 5.9 mg 2.64 348.27yl)-6-furan-2-ylmethyl-4,5,6,7- tetrahydro-2H-pyrazolo[3,4-c]pyridine 76-(4-Chlorobenzyl)-3-(5,6-dimethyl-1H- 4.7 mg 2.9 392.26benzoimidazol-2-yl)-4,5,6,7-tetrahydro- 2H-pyrazolo[3,4-c]pyridine 83-(5,6-Dimethyl-1H-benzoimidazol-2- 8.4 mg 2.71 364.24yl)-6-thiophen-2-ylmethyl-4,5,6,7- tetrahydro-2H-pyrazolo[3,4-c]pyridine9 3-(5,6-Dimethyl-1H-benzoimidazol-2- 11.7 mg 2.55 359.29yl)-6-pyridin-3-ylmethyl-4,5,6,7- tetrahydro-2H-pyrazolo[3,4-c]pyridine10 3-(5,6-Dimethyl-1H-benzoimidazol-2- 3.7 mg 2.72 338.32yl)-6-(2,2-dimethylpropyl)-4,5,6,7-tetrahydro-2H-pyrazolo[3,4-c]pyridine 113-(5,6-Dimethyl-1H-benzoimidazol-2- 5 mg 2.55 296.27yl)-6-ethyl-4,5,6,7-tetrahydro-2H- pyrazolo[3,4-c]pyridine 123-(5,6-Dimethyl-1H-benzoimidazol-2- 5.9 mg 2.76 338.3yl)-6-(3-methylbutyl)-4,5,6,7-tetrahydro- 2H-pyrazolo[3,4-c]pyridine 133-(5,6-Dimethyl-1H-benzoimidazol-2- 6.4 mg 2.62 310.29yl)-6-propyl-4,5,6,7-tetrahydro-2H- pyrazolo[3,4-c]pyridine 143-(5,6-Dimethyl-1H-benzoimidazol-2- 4.2 mg 2.97 386.31yl)-6-(3-phenylpropyl)-4,5,6,7- tetrahydro-2H-pyrazolo[3,4-c]pyridine 156-Butyl-3-(5,6-dimethyl-1H-benzo- 4.5 mg 2.68 324.28imidazol-2-yl)-4,5,6,7-tetrahydro-2H- pyrazolo[3,4-c]pyridine 166-Cyclopropylmethyl-3-(5,6-dimethyl- 3.9 mg 2.62 322.271H-benzoimidazol-2-yl)-4,5,6,7- tetrahydro-2H-pyrazolo[3,4-c]pyridine

EXAMPLE 10 Preparation of a Urea Library

The urea library can be prepared in the following way:

120 mg of3-(5,6-dimethyl-1H-benzimidazol-2-yl)-4,5,6,7-tetrahydro-2H-pyrazolo[3,4-c]pyridinehydrochloride are suspended in 3.6 ml of tetrahydrofuran in the presenceof 190 μl of triethylamine, and the suspension obtained is distributedinto each of the 9 test tubes, at a rate of 300 μl per tube.

The 9 isocyanates (Table 10) are weighed and added to each of the 9 testtubes.

TABLE 10 Isocyanates used Entry Name Amount 1 PHENYL ISOCYANATE 6.7 mg 2BENZYL ISOCYANATE 7.5 mg 3 2-CHLOROPHENYL ISOCYANATE 8.6 mg 43-CHLOROPHENYL ISOCYANATE 8.6 mg 5 4-CHLOROPHENYL ISOCYANATE 8.6 mg 6N-BUTYL ISOCYANATE 5.6 mg 7 2-THIENYL ISOCYANATE   7 mg 82-METHOXYPHENYL ISOCYANATE 8.4 mg 9 O-TOLYL ISOCYANATE 7.5 mg

The 9 reaction mixtures are shaken by means of orbital shaking atambient temperature for 2 hours, and are then evaporated to dryness.

Evaporation residues are each solubilized in 1 ml of DMSO and, for eachsolution obtained, a 10 μl sample is taken and diluted in 40 μl of DMSO(Gilson Liquid Handler Quad-X 215). Each sample in solution in DMSO thusobtained is analysed by LC/MS (method A).

The residual solutions are purified by LC/MS (method B). Afterpurification by LC/MS, the fractions containing the desired compoundsare (optionally combined) either evaporated to dryness (entries 1, 3, 6,8 and 9) or loaded onto SCX phase (500 mg of CUBCX1-HL phase; entries 2,4, 5 and 7). The SCX phases are subsequently washed with methanol andthen extracted with a solution of 2M ammonia in methanol. The extractionsolutions are collected in tared glass tubes, evaporated to dryness(Savant AES 2000 or Genevac HT8 centrifugal evaporator), weighed(Mettler Toledo Automated Workstation LA200) and diluted to 10 mM inDMSO (Gilson Liquid Handler Quad-Z 215). Each solution obtained isanalysed by LC/MS (method A).

The following compounds (Table 11) were isolated and characterized bymeans of their retention time and molecular peak in mass spectrometry(method A).

TABLE 11 Urea library obtained Amount of Retention Molecular producttime ion Entry Name obtained (minutes) detected 13-(5,6-Dimethyl-1H-benzoimidazol-2- 14.6 mg 3.04 387.28yl)-2,4,5,7-tetrahydropyrazolo[3,4-c]- pyridine-6-carboxylic acidphenyl- amide bistrifluoroacetate 2 3-(5,6-Dimethyl-1H-benzoimidazol-2-1.8 mg 2.78 401.29 yl)-2,4,5,7-tetrahydropyrazolo-[3,4-c]pyridine-6-carboxylic acid benzylamide 33-(5,6-Dimethyl-1H-benzoimidazol-2- 16 mg 2.92 421.25yl)-2,4,5,7-tetrahydropyrazolo- [3,4-c]pyridine-6-carboxylic acid(2-chlorophenyl)amide bistrifluoro- acetate 43-(5,6-Dimethyl-1H-benzoimidazol-2- 7.9 mg 3.89 421.24yl)-2,4,5,7-tetrahydropyrazolo- [3,4-c]pyridine-6-carboxylic acid(3-chlorophenyl)amide 5 3-(5,6-Dimethyl-1H-benzoimidazol-2- 9.8 mg 3.36421.25 yl)-2,4,5,7-tetrahydropyrazolo- [3,4-c]pyridine-6-carboxylic acid(4-chlorophenyl)amide 6 3-(5,6-Dimethyl-1H-benzoimidazol-2- 2.4 mg 2.8367.31 yl)-2,4,5,7-tetrahydropyrazolo- [3,4-c]pyridine-6-carboxylic acidbutylamide bistrifluoroacetate 7 3-(5,6-Dimethyl-1H-benzoimidazol-2- 3.8mg 2.77 393.24 yl)-2,4,5,7-tetrahydropyrazolo-[3,4-c]pyridine-6-carboxylic acid thiophen-2-ylamide 83-(5,6-Dimethyl-1H-benzoimidazol-2- 14.4 mg 3.14 417.28yl)-2,4,5,7-tetrahydropyrazolo- [3,4-c]pyridine-6-carboxylic acid(2-methoxyphenyl)amide bistrifluoroacetate 93-(5,6-Dimethyl-1H-benzoimidazol-2- 16.2 mg 2.68 401.29yl)-2,4,5,7-tetrahydropyrazolo- [3,4-c]pyridine-6-carboxylic acido-tolylamide bistrifluoroacetate

EXAMPLE 113-(5,6-Dimethyl-1H-benzimidazol-2-yl)-4,5,6,7-tetrahydro-2H-pyrazolo[3,4-c]pyridine

3-(5,6-Dimethyl-1H-benzimidazol-2-yl)-4,5,6,7-tetrahydro-2H-pyrazolo-[3,4-c]pyridinecan be prepared in the following way:

9 ml of water and 2.8 ml of trifluoroacetic acid are added to a solutionof 670 mg of tert-butyl3-(2-amino-4,5-dimethylphenylcarbamoyl)-2,4,5,7-tetrahydropyrazolo[3,4-c]pyridine-6-carboxylatein 9 ml of THF. After stirring for 2 hours at 80° C., the reactionmedium is concentrated under reduced pressure. It is then taken up inwater and the precipitate formed is recovered by filtration throughsintered glass, washed with an aqueous 1N sodium hydroxide solution anddried. The aqueous phase obtained is subsequently extracted withdichloromethane and the organic phase is then dried over magnesiumsulphate and concentrated under reduced pressure. The residue obtainedand the precipitate are combined and then solubilized in methanol with afew drops of DMF. This solution is then loaded onto MEGA BE-SCX phase.The SCX phase is subsequently washed with methanol and extracted with asolution of 2M ammonia in methanol. The extraction solution obtained isthen concentrated under reduced pressure.

46 mg of3-(5,6-dimethyl-1H-benzimidazol-2-yl)-4,5,6,7-tetrahydro-2H-pyrazolo[3,4-c]pyridineare thus obtained in the form of a beige powder, the characteristics ofwhich are as follows:

EI: m/z = 267 M^(+.) base peak m/z = 238 [M − NHCH₂]⁺ m/z = 209 [M −C₃H₈N]^(+.)

¹H NMR (300 MHz, (CD₃)₂SO, δ in ppm): 2.31 and 2.32 (2 s: 6H in total);2.81 (broad t, J=5 Hz: 2H); 2.92 (broad t, J=5 Hz: 2H); 3.83 (broad s:2H); 7.22 (broad s: 1H); 7.40 (broad s: 1H); 12.28 (unresolved peak:1H); 12.73 (unresolved peak: 1H).

EXAMPLE 123-(5,6-Dimethyl-1H-benzimidazol-2-yl)-4,5,6,7-tetrahydro-2H-pyrazolo[3,4-c]pyridinehydrochloride

3-(5,6-Dimethyl-1H-benzimidazol-2-yl)-4,5,6,7-tetrahydro-2H-pyrazolo[3,4-c]pyridinecan be prepared in the following way:

9 ml of an aqueous 5N hydrochloric acid solution are added to a solutionof 1.7 g of tert-butyl3-(2-amino-4,5-dimethylphenylcarbamoyl)-2,4,5,7-tetrahydropyrazolo[3,4-c]pyridine-6-carboxylatein 40 ml of ethanol. After stirring at 80° C. for 60 hours, the reactionmedium is brought back to ambient temperature. The precipitate formed isrecovered by filtration through sintered glass and dried. 1.04 g of3-(5,6-dimethyl-1H-benzimidazol-2-yl)-4,5,6,7-tetrahydro-2H-pyrazolo[3,4-c]pyridinehydrochloride are thus obtained in the form of a beige powder, thecharacteristics of which are as follows:

EI: m/z = 267 M^(+.) base peak m/z = 238 [M − CH₂NH]⁺ m/z = 209 [M −C₃H₈N]^(+.) m/z = 36 [HCl]⁺

¹H NMR (300 MHz, (CD₃)₂SO with addition of a few drops of CD₃COOD, δ inppm): 2.40 (s: 6H); 3.23 (broad t, J=5.5 Hz: 2H); 3.45 (t, J=5.5 Hz:2H); 4.45 (s: 2H); 7.54 (s: 2H).

tert-Butyl3-(2-amino-4,5-dimethylphenylcarbamoyl)-2,4,5,7-tetrahydro-pyrazolo[3,4-c]pyridine-6-carboxylatecan be prepared in the following way: 8.5 g of HBTU and also 2.9 g ofdiisopropylethylamine are added, at ambient temperature, to a solutionof 3 g of (tert-butyl2,4,5,7-tetrahydropyrazolo-[3,4-c]pyridine-6-carboxylate)-3-carboxylicacid in 50 ml of anhydrous DMF. After twenty minutes' stirring atambient temperature, 3.06 g of 4,5-diamino-o-xylene are added. Afterstirring at ambient temperature for 60 hours, the reaction medium isdiluted in 3 l of an aqueous NaHCO₃ solution at pH greater than 7,containing 20 g of NaCl. The aqueous phase is extracted three times with1 l of ethyl acetate, and the combined organic phases are then driedover magnesium sulphate and concentrated under reduced pressure. Thecrude residue obtained is taken up in 150 ml of dichloromethane and theinsoluble material is removed by filtration through sintered glass. Thefiltrate is then concentrated under reduced pressure and purified bychromatography on silica (20–45 μm Amicon) with a gradient of from 50 to100% of ethyl acetate in cyclohexane. The fractions containing thedesired product are combined and concentrated under reduced pressure.4.37 g of tert-butyl3-(2-aminophenylcarbamoyl)-2,4,5,7-tetrahydro-pyrazolo[3,4-c]pyridine-6-carboxylateare thus obtained in the form of a beige powder, the characteristics ofwhich are as follows:

EI: m/z = 385 M^(+.) base peak m/z = 329 [M − C₄H₈]^(+.) m/z = 312 [M −C₄H₉O]⁺ m/z = 57 [C₄H₉]⁺

¹H NMR (300 MHz, (CD₃)₂SO, δ in ppm): 1.46 (s: 9H); 2.10 and 2.12 (2 s:3each); 2.77 (mt: 2H); 3.58 (t, J=5.5 Hz: 2H); 4.53 (s: 2H); 4.57(unresolved peak: 2H); 6.60 (s: 1H); 7.14 (broad s: 1H); 9.10(unresolved peak: 1H); 13.08 (unresolved peak: 1H).

EXAMPLE 133-(1H-Benzimidazol-2-yl)-4,5,6,7-tetrahydro-2H-pyrazolo-[3,4-c]pyridineHydrochloride

3-(1H-Benzimidazol-2-yl)-4,5,6,7-tetrahydro-2H-pyrazolo[3,4-c]pyridinehydrochloride can be prepared in the following way:

2.2 ml of an aqueous 5N hydrochloric acid solution are added to asolution of 200 mg of tert-butyl3-(2-aminophenylcarbamoyl)-2,4,5,7-tetrahydro-pyrazolo[3,4-c]pyridine-6-carboxylatein 1 ml of ethanol. After stirring for 20 hours at 80° C., the reactionmedium is brought back to ambient temperature. The insoluble material isremoved by filtration through sintered glass and the filtrate isconcentrated under reduced pressure. 84 mg of3-(1H-benzimidazol-2-yl)-4,5,6,7-tetrahydro-2H-pyrazolo[3,4-c]pyridinehydrochloride are thus obtained in the form of an orange-colouredpowder, the characteristics of which are as follows:

LC/MS (method A): molecular ion detected: 240.26; retention time=1.68minutes

tert-Butyl3-(2-aminophenylcarbamoyl)-2,4,5,7-tetrahydro-pyrazolo[3,4-c]-pyridine-6-carboxylatecan be prepared in the following way:

425 mg of HBTU and also 145 mg of diisopropylethylamine are added, atambient temperature, to a solution of 150 mg of (tert-butyl2,4,5,7-tetrahydro-pyrazolo[3,4-c]pyridine-6-carboxylate)-3-carboxylicacid in 1 ml of anhydrous DMF. After stirring for twenty minutes atambient temperature, 121 mg of orthophenylenediamine are added.

After stirring at ambient temperature for 20 hours, the reaction mediumis diluted in 100 ml of water and 50 ml of ethyl acetate. The aqueousphase is extracted three times with 50 ml of ethyl acetate and thecombined organic phases are then dried over magnesium sulphate andconcentrated under reduced pressure. The crude residue obtained ispurified by HPLC (reverse phase C18 Lichroprep 12 μm) with a lineargradient of from 5 to 95% of acetonitrile comprising 0.07% (v/v) oftrifluoroacetic acid in water comprising 0.07% (v/v) of trifluoroaceticacid, at a flow rate of 10 ml/min. The fractions containing the desiredproduct are combined and loaded onto MEGA BE-SCX phase. The SCX phase issubsequently washed with methanol and extracted with a solution of 2Mammonia in methanol. The extraction solution obtained is thenconcentrated under reduced pressure. 200 mg of tert-butyl3-(2-aminophenylcarbamoyl)-2,4,5,7-tetrahydropyrazolo[3,4-c]pyridine-6-carboxylateare thus obtained in the form of a beige powder, the characteristics ofwhich are as follows:

LC/MS (method A): molecular ion detected: 358.34; retention time=3.19minutes.

EXAMPLE 14 Preparation of a Sulphonamide Library

The sulphonamide library can be prepared in the following way:

40 mg of6-[3-(5,6-dimethyl-1H-benzimidazol-2-yl)-2,4,5,7-tetrahydro-pyrazolo[3,4-c]pyridin-6-yl]pyridin-3-ylamineare suspended in 2 ml of dichloromethane and the solution obtained isdistributed into 4 test tubes, at a rate of 500 μl per tube.

The 4 sulphonyl chlorides (Table 12) are weighed and added to each ofthe 4 test tubes, followed by 15.6 μl of triethylamine.

TABLE 12 Sulphonyl chlorides used Entry Name Amount 1THIOPHENE-2-SULPHONYL CHLORIDE 5.6 mg 2 4-METHOXYBENZENESULPHONYLCHLORIDE 6.3 mg 3 2-CHLOROBENZENESULPHONYL CHLORIDE 6.4 mg 42-METHOXY-4-METHYLBENZENESULPHONYL 6.8 mg CHLORIDE

The 4 reaction mixtures are shaken by means of orbital shaking at 40° C.for 15 h.

For each reaction mixture, a 5 μl sample is taken and diluted in 100 μlof DMSO (Gilson Liquid Handler Quad-Z 215). Each sample in solution inDMSO thus obtained is analysed by LC/MS (method A).

The 4 reaction mixtures are then evaporated to dryness and theevaporation residues are each solubilized in 500 μl of DMSO and thesolutions obtained are purified by LC/MS (method B). After purificationby LC/MS, the fractions containing the desired compounds are (optionallycombined) loaded onto SCX phase (500 mg of CUBCX1-HL phase). The SCXphases are subsequently washed with methanol and then extracted with asolution of 2M ammonia in methanol. The extraction solutions arecollected in tared glass tubes, evaporated to dryness (Savant AES 2000or Genevac HT8 centrifugal evaporator), weighed (Mettler ToledoAutomated Workstation LA200) and diluted to 10 mM in DMSO (Gilson LiquidHandler Quad-Z 215). Each solution obtained is analysed by LC/MS (methodA).

The following compounds (Table 13) were isolated and characterized bymeans of their retention time and molecular peak in mass spectrometry(method A)

TABLE 13 Sulphonamide library obtained Amount Retention Molecular ofproduct time ion Entry Name obtained (minutes) detected 1N-{6-[3-(5,6-Dimethyl-1H- 2.9 mg 3.07 506.21 benzimidazol-2-yl)-2,4,5,7-tetrahydropyrazolo[3,4-c]pyridin- 6-yl]pyridin-3-yl}thiophene-2-sulphonamide 2 N-{6-[3-(5,6-Dimethyl-1H- 3.0 mg 3.20 530.25benzimidazol-2-yl)-2,4,5,7- tetrahydropyrazolo[3,4-c]pyridin-6-yl]pyridin-3-yl}-4-methoxy- benzenesulphonamide 32-Chloro-N-{6-[3-(5,6-dimethyl- 3.0 mg 3.38 534.211H-benzimidazol-2-yl)-2,4,5,7- tetrahydropyrazolo[3,4-c]pyridin-6-yl]pyridin-3-yl}benzene- sulphonamide 4 N-{6-[3-(5,6-Dimethyl-1H- 3.8mg 3.38 544.26 benzimidazol-2-yl)-2,4,5,7-tetrahydropyrazolo[3,4-c]pyridin- 6-yl]pyridin-3-yl}-2-methoxy-4-methylbenzenesulphonamide

EXAMPLE 156-[3-(5,6-Dimethyl-1H-benzimidazol-2-yl)-2,4,5,7-tetrahydro-pyrazolo[3,4-c]pyridin-6-yl]pyridin-3-ylamine

6-[3-(5,6-Dimethyl-1H-benzimidazol-2-yl)-2,4,5,7-tetrahydropyrazolo-[3,4-c]pyridin-6-yl]pyridin-3-ylaminecan be prepared in the following way:

55 mg of Pd/CaCO₃ 10% are added to a solution of 545 mg of3-(5,6-dimethyl-1H-benzimidazol-2-yl)-6-(5-nitropyridin-2-yl)-4,5,6,7-tetrahydro-2H-pyrazolo[3,4-c]pyridinein 60 ml of ethanol. After stirring for 15 hours at 35° C. under 3 barof hydrogen, the reaction medium is brought back to ambient temperature,filtered through celite and then concentrated under reduced pressure.300 mg of6-[3-(5,6-dimethyl-1H-benzimidazol-2-yl)-2,4,5,7-tetrahydropyrazolo[3,4-c]pyridin-6-yl]-pyridin-3-ylamine are thus obtained in the form ofa brown powder, the characteristics of which are as follows:

EI m/z = 359 M⁺ base peak m/z = 266 (M—C₅H₅N₂)⁺

EXAMPLE 163-(5,6-Dimethyl-1H-benzimidazol-2-yl)-6-(5-nitropyridin-2-yl)-4,5,6,7-tetrahydro-2H-pyrazolo[3,4-c]pyridine

3-(5,6-Dimethyl-1H-benzimidazol-2-yl)-6-(5-nitropyridin-2-yl)-4,5,6,7-tetrahydro-2H-pyrazolo[3,4-c]pyridinecan be prepared in the following way:

287 mg of 2-chloro-5-nitropyridine and 500 mg of potassium carbonate areadded to a solution of 500 mg of3-(5,6-dimethyl-1H-benzimidazol-2-yl)-4,5,6,7-tetrahydro-2H-pyrazolo[3,4-c]pyridinehydrochloride in 5 ml of dimethylformamide. After stirring for 20 hoursat ambient temperature, the reaction medium is added to 50 ml of water.The precipitate formed is recovered by filtration through sinteredglass, washed with 3 times 15 ml of water and then dried under reducedpressure. 548 mg of3-(5,6-dimethyl-1H-benzimidazol-2-yl)-6-(5-nitropyridin-2-yl)-4,5,6,7-tetrahydro-2H-pyrazolo-[3,4-c]pyridineare thus obtained in the form of a yellow powder, the characteristics ofwhich are as follows:

EI: m/z = 389 M^(+.) base peak m/z = 266 (M − C₅H₃N₂O₂)⁺

EXAMPLE 176-{5-[3-(2-Fluoro-5-trifluoromethylphenyl)ureido]pyridin-2-yl}-4,5,6,7-tetrahydro-2H-pyrazolo[3,4-c]pyridine-3-carboxamide

6-{5-[3-(2-Fluoro-5-trifluoromethylphenyl)ureido]pyridin-2-yl}-4,5,6,7-tetrahydro-2H-pyrazolo[3,4-c]pyridine-3-carboxamidecan be prepared in the following way from ethyl6-(5-tert-butoxycarbonylaminopyridin-2-yl)-4,5,6,7-tetrahydro-2H-pyrazolo[3,4-c]pyridine-3-carboxylate:

The ethyl ester of ethyl6-(5-tert-butoxycarbonylaminopyridin-2-yl)-4,5,6,7-tetrahydro-2H-pyrazolo[3,4-c]pyridine-3-carboxylateis converted to carboxamide by amidation using a solution of aqueousammonia, and results in the obtaining of6-(5-tert-butoxycarbonylaminopyridin-2-yl)-4,5,6,7-tetrahydro-2H-pyrazolo[3,4-c]pyridine-3-carboxamide.

EI: m/z=358

The amine group of the6-(5-tert-butoxycarbonylaminopyridin-2-yl)-4,5,6,7-tetrahydro-2H-pyrazolo[3,4-c]pyridine-3-carboxamideis deprotected in acid medium (trifluoroacetic acid in dichloromethane)and results in the obtaining of6-(5-aminopyridin-2-yl)-4,5,6,7-tetrahydro-2H-pyrazolo[3,4-c]pyridine-3-carboxamide.

EI: m/z=258

The urea function is introduced onto the6-(5-aminopyridin-2-yl)-4,5,6,7-tetrahydro-2H-pyrazolo[3,4-c]pyridine-3-carboxamideaccording to the method described in Example 1, using2-fluoro-5-(trifluoromethyl)phenyl isocyanate, and results in theobtaining of6-{5-[3-(2-fluoro-5-trifluoromethylphenyl)-ureido]pyridin-2-yl}4,5,6,7-tetrahydro-2H-pyrazolo[3,4-c]pyridine-3-carb-oxamide.

EI: m/z=463

EXAMPLE 18 Ethyl6-(5-tert-butoxycarbonylaminopyridin-2-yl)-4,5,6,7-tetra-hydro-2H-pyrazolo[3,4-c]pyridine-3-carboxylate

Ethyl6-(5-tert-butoxycarbonylaminopyridin-2-yl)-4,5,6,7-tetrahydro-2H-pyrazolo[3,4-c]pyridine-3-carboxylatecan be prepared in the following way:

5 mg of Pd/C 10% and 38 mg of di-tert-butyl dicarbonate are added to asolution of 50 mg of ethyl6-(5-nitropyridin-2-yl)-4,5,6,7-tetrahydro-2H-pyrazolo[3,4-c]pyridine-3-carboxylatein 6 ml of methanol. After stirring for 12 hours at ambient temperatureunder 3 bar of hydrogen, the reaction medium is filtered through celiteand then concentrated under reduced pressure. The reaction crudeobtained is purified by flash chromatography (SiO₂, CH₂Cl₂/MeOH gradient75/25 to 25/75). 20 mg of ethyl6-(5-tert-butoxycarbonylaminopyridin-2-yl)-4,5,6,7-tetrahydro-2H-pyrazolo[3,4-c]pyridine-3-carboxylateare thus obtained in the form of a white powder, the characteristics ofwhich are as follows:

EI: m/z = 387 M^(+.) m/z = 331 (M − C₄H₈)^(+.) base peak m/z = 286 (m/z= 331 − CO₂H)⁺ m/z = 194 C₉H₁₂N₃O₂ ⁺ m/z = 57 C₄H₉ ⁺

EXAMPLE 19 Ethyl 6-(5-nitropyridin-2-yl)-4,5,6,7-tetrahydro-2H-pyrazolo[3,4-c]pyridine-3-carboxylate

Ethyl6-(5-nitropyridin-2-yl)-4,5,6,7-tetrahydro-2H-pyrazolo[3,4-c]pyridine-3-carboxylatecan be prepared in the following way:

522 mg of 2-chloro-5-nitropyridine are added to a solution of 1 g of3-(5,6-dimethyl-1H-benzimidazol-2-yl)-4,5,6,7-tetrahydro-2H-pyrazolo[3,4-c]pyridinetrifluoroacetate in 10 ml of pyridine. After stirring for 20 hours atambient temperature, the reaction medium is concentrated under reducedpressure. The precipitate formed is recovered by filtration throughsintered glass, washed with 3 times 15 ml of water, and dried underreduced pressure. The reaction crude obtained is purified by flashchromatography (SiO₂, cyclohexane/EtOAc gradient 75/25 to 25/75). 450 mgof ethyl6-(5-nitro-pyridin-2-yl)-4,5,6,7-tetrahydro-2H-pyrazolo[3,4-c]pyridine-3-carboxylateare thus obtained in the form of a yellow powder, the characteristics ofwhich are as follows:

EI: m/z = 317 M^(+.) base peak m/z = 271 (M − NO₂)^(+.) m/z = 194 (M −C₅H₃N₂O₂)⁺ m/z = 148 (m/z = 194 − C₂H₆O)⁺

EXAMPLE 20 Ethyl4,5,6,7-tetrahydro-2H-pyrazolo[3,4-c]pyridine-3-carboxylatetrifluoroacetate

Ethyl 4,5,6,7-tetrahydro-2H-pyrazolo[3,4-c]pyridine-3-carboxylatetrifluoro-acetate can be prepared in the following way:

50 ml of water followed by 12 ml of trifluoroacetic acid are added to asolution of 3 g of 6-tert-butyl-3-ethyl2,4,5,7-tetrahydropyrazolo[3,4-c]pyridyl-3,6-dicarboxylate in 50 ml oftetrahydrofuran. After stirring for 2 hours at reflux, the reactionmedium is brought back to ambient temperature and a saturated aqueousNa₂CO₃ solution is added until a basic pH is obtained. The aqueous phaseobtained is extracted 3 times with ethyl acetate. The combined organicphases are dried over magnesium sulphate and concentrated under reducedpressure. 1.49 g of ethyl4,5,6,7-tetrahydro-2H-pyrazolo[3,4-c]pyridine-3-carboxylatetrifluoroacetate are thus obtained, the characteristics of which are asfollows:

EI: m/z = 195 M^(+.) m/z = 166 (M − CH₃N)^(+.) base peak m/z = 138 (M −C₃H₇N)^(+.) m/z = 120 (m/z = 166 − C₂H₆O)^(+.) m/z = 92 (m/z = 120 −CO)^(+.)Measurements of the Inhibitory Potential of the Products with Respect tothe Activity of the Tie2 and KDR Kinases:

The inhibitory activity of the products with respect to the Tie2 and KDRkinases is tested according to the experimental protocols describedbelow.

1. Tie2

The coding sequence of human Tie2 corresponding to amino acids 776–1124of the intracellular domain was generated by PCR using the cDNA isolatedfrom human placenta as model. This sequence was introduced into abaculovirus expression vector pFastBacGT in the form of a GST fusionprotein.

The inhibitory effect of the molecules is determined in an assay forphosphorylation of PLC by Tie2 in the presence of GST-Tie2 purified toapproximately 80% homogeneity. The substrate is made up of the SH2-SH3fragments of PLC, the latter being expressed in the form of a GST fusionprotein.

The kinase activity of Tie2 is measured in a 20 mM MOPS buffer, pH 7.2,containing 10 mM MgCl₂, 10 mM MnCl₂, 1 mM DTT and 10 mM ofglycerophosphate. A reaction mixture made up of 70 μl of kinase buffercontaining 100 ng of GST-Tie2 enzyme per well is placed in a flashplate96-well plate kept on ice. 10 μl of the molecule to be tested, dilutedin DMSO, at a concentration of at most 10% are then added. For a givenconcentration, each measurement is carried out in quadruplicate. Thereaction is initiated by adding 20 μl of solution containing 2 μg ofGST-PLC, 2 μM of cold ATP and 1 μCi of ³³P[ATP]. After incubation for 1hour at 37° C., the reaction is stopped by adding 1 volume (100 μl) of200 mM EDTA. After removal of the incubation buffer, the wells arewashed three times with 300 μl of PBS. The radioactivity is measured ona Wallac MicroBeta 1450.

The inhibition of the Tie2 activity is calculated and expressed aspercentage inhibition with respect to the control activity determined inthe absence of compound.

2. KDR

The inhibitory effect of the compounds is determined in an assay forphosphorylation of substrate by the KDR enzyme in vitro using ascintillation technique (96-well plate, NEN).

The cytoplasmic domain of the human KDR enzyme was cloned in GST fusionform into the baculovirus expression vector pFastBac. The protein wasexpressed in SF21 cells and purified to approximately 60% homogeneity.

The KDR kinase activity is measured in 20 mM MOPS, 10 mM MgCl₂, 10 mMMnCl₂, 1 mM DTT, 2.5 mM EGTA, 10 mM b-glycerophosphate, pH=7.2, in thepresence of 10 mM MgCl₂, 100 μM Na₃VO₄ and 1 mM NaF. 10 μl of thecompound are added to 70 μl of kinase buffer containing 100 ng of KDRenzyme at 4° C. The reaction is initiated by adding 20 μl of solutioncontaining 2 μg of substrate (SH2-SH3 fragment of PLCγ expressed in theform of an GST fusion protein), 2 μCi γ³³P[ATP] and 2 μM cold ATP. Afterincubation for 1 hour at 37° C., the reaction is stopped by adding 1volume (100 μl) of 200 mM EDTA. The incubation buffer is removed and thewells are washed three times with 300 μl of PBS. The radioactivity ismeasured in each well using a Top Count NXT (Packard) radioactivitycounter.

The background noise is determined by measuring the radioactivity infour different wells containing the radioactive ATP and the substratealone.

A total activity control is measured in four different wells containingall the reagents (γ³³P-[ATP], KDR and PLC-γ substrate) but in theabsence of compound.

The inhibition of the KDR activity with the compound of the invention isexpressed as percentage inhibition of the control activity determined inthe absence of compound.

The compound SU5614 (Calbiochem) (1 μM) is included in each plate asinhibition control.

Results:

Tie2 KDR % Inhib at 10 μM (FRX) % Inhib at 10 μM Chemistry Assay 1 Assay2 Assay 1 Assay 2 P-31378-112-3 78.7 79.1 59.0 56.5

P-31378-112-6 92.7 93.8 71.4 68.0

P-31378-112-2 92.6 92.6 98.1 97.4

P-31378-112-8^(H) 87.8 93.3 86.8 89.0

P-31378-112-4 83.1 87.2 56.2 50.1

P-31378-112-9 82.6 85.9 53.2 45.1

P-31378-112-14 69.7 73.8 19.5 13.0

P-31378-112-1 83.1 88.1 96.5 96.7

P-31378-112-16 88.7 88.3 90.2 89.1

P-31378-112-5 87.1 86.4 94.7 95.4

P-31378-112-10 90.6 87.9 42.3 28.7

P-31378-112-7 92.7 92.0 68.6 64.3

P-31378-112-15 80.1 83.8 75.5 77.4

KDR % Inhib. Tie2 % Inhib. Chemistry 10 μM 10 μM

77.20 Chemistry 2

78.10 Chemistry 3

25.40 Chemistry 4

88.80 Chemistry 5

2.10 41.2 Chemistry 6

12.15 48.6 Chemistry 7

85.45 64.3 Chemistry 8

15.45 56.7 Chemistry 9

79.55 83.9 Chemistry 10

70.35 66.1 Chemistry 11

69.05 60.3 Chemistry 12

84.50 41.5 Chemistry 13

14.85 50.5 Chemistry 14

39.30 69.4 Chemistry 15

−5.75 36.4 Chemistry 16

−2.90 50.7 Chemistry 17

−3.70 55.0 Chemistry 18

17.90 68.1 Chemistry 19

1.65 47.6 Chemistry 20

1.85 33.0 Chemistry 21

3.65 24.9 Chemistry 22

48 92.5 Chemistry 23

70.75 91.5 Chemistry 24

31.85 89.1 Chemistry 25

27.75 89.1 Chemistry 26

22.95 65.4 Chemistry 27

16.50 78.5 Chemistry 28

13.35 47.5 Chemistry 29

15.30 44.2 Chemistry 30

45.70 93.2 Chemistry 31

46.45 91.5 Chemistry 32

57.70 95.9 Chemistry 33

18.15 84.7 Chemistry 34

27.40 88.8 Chemistry 35

49.40 90.8 Chemistry 36

41.20 88.3 Chemistry 37

20.85 84.6 Chemistry 38

30.90 97.9 Chemistry 39

14.40 83.4 Chemistry 40

72.50 86.3 Chemistry 41

52.65 97.4 Chemistry 42

10.90 84.4 Chemistry 43

41.10 90.8 Chemistry 44

33.40 94.1 Chemistry 45

73.05 96.7 Chemistry 46

78.55 87.7 Chemistry 47

41.95 83.8 Chemistry 48

12.20 84.8 Chemistry 49

74.75 82.4 Chemistry 50

38.70 72.9 Chemistry 51

29.35 89.3 Chemistry 52

74.55 92.7 Chemistry 53

11.25 81.3 Chemistry 54

15.95 65.4 Chemistry 55

46.65 92.5 Chemistry 56

75.15 94.6 Chemistry 57

27.90 82.3 Chemistry 58

17.6 33.2 Chemistry 59

33.55 64.85 Chemistry 60

12.7 3.75 Chemistry 61

12.85 21 Chemistry 62

−0.3 23.75 Chemistry 63

18 38.5 Chemistry 64

63.3 85.9 Chemistry 65

8.1 18.05 Chemistry 67

19.5 19.1 Chemistry 68

18.55 59.3 Chemistry 69

92.1 65.5 Chemistry 70

98

100

99

100

1. A compound of formula (I), including its tautomers:

wherein: L is a bond; X is chosen from a bond, CH₂, CO, SO₂, CONH andCOO; R1 is OH or H, or is chosen from alkyl, cycloalkyl, heterocycle,aryl, heteroaryl, chosen from benzimidazolyl, indolyl or pyrrolyl, allof which is optionally substituted, and, when X is a bond, then R1 mayalso be halogen; R2 is H or is chosen from alkyl, alkylene, cycloalkyl,heterocycle, aryl, heteroaryl, chosen from pyridyl, furanyl or thienyl,all of which is optionally substituted; and in that the optionalsubstituents are chosen independently from R3, O—R3, halogen, NO₂,SO₂—R3, CO—R3, SO₂NH—R3, CONH—R3, N—(R3)₂, NHCO—R3, NHSO₂—R3, NHCONH—R3,NHSO₂NH—R3, OCO—R3, COO—R3, OSO₂—R3, SO₂O—R3, OCONH—R3 and OSO₂NH—R3,wherein R3 is H or is chosen independently from alkyl, cycloalkyl,alkenyl, aryl, heteroaryl chosen from benzimidazolyl, indolyl, pyridyl,furanyl, thienyl or pyrrolyl, heterocycle, all of which is optionallysubstituted with halogen, aryl, heteroaryl chosen from benzimidazolyl,indolyl, pyridyl, furanyl, thienyl or pyrrolyl, R4, OR4 or N(R4)₂,wherein each R4 is chosen independently from H, C₁–C₄ alkyl andhalogenated C₁–C₄ alkyl, or a racemate, a stereoisomer, an enantiomer,or a mixture in any combination thereof, or a pharmaceuticallyacceptable salt thereof; with the proviso that: when R1 is OH and X is abond or COO, then R2 is not hydrogen or methyl; when R1 is methyl,cyclopropyl or 4-fluorophenyl, and X is a bond, then R2 is not hydrogen.2. The compound as set forth in claim 1, which is of the formula (II):

and its tautomers, wherein: X is chosen from a bond, CH₂, CO, SO₂, CONHand COO; R1 is chosen from alkyl, cycloalkyl, heterocycle, aryl,heteroaryl chosen from benzimidazolyl, indolyl, or pyrrolyl, all ofwhich is optionally substituted; R2 is H or is chosen from alkyl,alkylene, cycloalkyl, heterocycle, aryl, heteroaryl chosen from pyridyl,furanyl or thienyl, all of which is optionally substituted; and in thatthe optional substituents are chosen independently from R3, O—R3,halogen, NO₂, SO₂—R3, CO—R3, SO₂NH—R3, CONH—R3, N—(R3)₂, NHCO—R3,NHSO₂—R3, NHCONH—R3, NHSO₂NH—R3, OCO—R3, COO—R3, OSO₂—R3, SO₂O—R3,OCONH—R3 and OSO₂NH—R3, wherein R3 is H or is chosen independently fromalkyl, cycloalkyl, alkenyl, aryl, heteroaryl, chosen frombenzimidazolyl, indolyl, pyridyl, furanyl, thienyl or pyrrolyl,heterocycle, all of which is optionally substituted with halogen, aryl,heteroaryl chosen from benzimidazolyl, indolyl, pyridyl, furanyl,thienyl or pyrrolyl, R4, OR4 or N(R4)₂, in which each R4 is chosenindependently from H and C₁–C₄ alkyl, or a racemate, a stereoisomer, anenantiomer, or a mixture in any combination thereof, or apharmaceutically acceptable salt thereof.
 3. The compound as set forthin claim 1, wherein R1 is heteroaryl chosen from benzimidazolyl, indolylor pyrrolyl, which is optionally substituted.
 4. The compound as setforth in claim 2, wherein R1 is heteroaryl chosen from benzimidazolyl,indolyl or pyrrolyl, which is optionally substituted.
 5. The compound asset forth in claim 3, wherein R1 is chosen from benzimidazolyl, indolyl,pyrrolyl, optionally substituted with halogen, R4 or O—R4, wherein R4 ischosen independently from H and C₁–C₄ alkyl.
 6. The compound as setforth in claim 5, wherein R1 is chosen from benzimidazol-2-yl,indol-2-yl, pyrrol-2-yl, optionally substituted with halogen, R4 orO—R4, wherein R4 is chosen independently from H and C₁–C₄ alkyl.
 7. Thecompound as set forth in claim 1, wherein R2 is chosen from phenyl,pyridyl, thienyl, C₁–C₄ alkyl, and C₃–C₇ cycloalkyl, all of which isoptionally substituted.
 8. The compound as set forth in claim 1, whereinX is chosen from CO and SO₂.
 9. The compound as set forth in claim 1,wherein R1 is H.
 10. The compound as set forth in claim 1, wherein R1 issubstituted aryl.
 11. The compound as set forth in claim 1, wherein X isa bond, and R2 is chosen from substituted aryl and substitutedheteroaryl wherein heteroaryl is chosen from pyridyl or thienyl.
 12. Thecompound as set forth in claim 1, which is:


13. A pharmaceutical composition comprising a compound of formula (I),including its tautomers, in combination with one or morepharmaceutically acceptable excipient, diluent or a carrier:

wherein: L is a bond; X is chosen from a bond, CH₂, CO, SO₂, CONH andCOO; R1 is OH or H, or is chosen from alkyl, cycloalkyl, heterocycle,aryl, heteroaryl, chosen from benzimidazolyl, indolyl or pyrrolyl, allof which is optionally substituted, and, when X is a bond, then R1 mayalso be halogen; R2 is H or is chosen from alkyl, alkylene, cycloalkyl,heterocycle, aryl, heteroaryl chosen from pyridyl, furanyl or thienyl,all of which is optionally substituted; and in that the optionalsubstituents are chosen independently from R3, O—R3, halogen, NO₂,SO₂—R3, CO—R3, SO₂NH—R3, CONH—R3, N—(R3)₂, NHCO—R3, NHSO₂—R3, NHCONH—R3,NHSO₂NH—R3, OCO—R3, COO—R3, OSO₂—R3, SO₂O—R3, OCONH—R3 and OSO₂NH—R3,wherein R3 is H or is chosen independently from alkyl, cycloalkyl,alkenyl, aryl, heteroaryl, chosen from benzimidazolyl, indolyl, pyridyl,furanyl, thienyl or pyrrolyl, heterocycle, all of which is optionallysubstituted with halogen, aryl, heteroaryl chosen from benzimidazolyl,indolyl, pyridyl, furanyl, thienyl or pyrrolyl, R4, OR4 or N(R4)₂,wherein each R4 is chosen independently from H, C₁–C₄ alkyl andhalogenated C₁–C₄ alkyl, or a racemate, a stereoisomer, an enantiomer,or a mixture in any combination thereof, or a pharmaceuticallyacceptable salt thereof; with the proviso that: when R1 is OH and X is abond or COO, then R2 is not hydrogen or methyl; when R1 is methyl,cyclopropyl or 4-fluorophenyl, and X is a bond, then R2 is not hydrogen.